U.S. patent application number 11/380291 was filed with the patent office on 2006-12-21 for method for improving pharmacokinetics of protease inhibitors and protease inhibitor precursors.
This patent application is currently assigned to AMBRILIA BIOPHARMA INC.. Invention is credited to Michael Ge, Guy Milot, Chandra Panchal, Marco Petrella, Brent Richard Stranix, Jinzi J. Wu.
Application Number | 20060287316 11/380291 |
Document ID | / |
Family ID | 37214407 |
Filed Date | 2006-12-21 |
United States Patent
Application |
20060287316 |
Kind Code |
A1 |
Wu; Jinzi J. ; et
al. |
December 21, 2006 |
METHOD FOR IMPROVING PHARMACOKINETICS OF PROTEASE INHIBITORS AND
PROTEASE INHIBITOR PRECURSORS
Abstract
The present invention provides methods for improving the
pharmacokinetics of protease inhibitors and protease inhibitor
precursors and pharmaceutical composition comprising protease
inhibitors or protease inhibitor precursors of formula I and a
cytochrome P450 monooxigenase inhibitor; ##STR1## when the compound
of formula I comprises an amino group, pharmaceutically acceptable
ammonium salts thereof, wherein R.sub.1 may be, for example,
(HO).sub.2P(O)--, (NaO).sub.2P(O)--, alkyl-CO-- or cycloalkyl-CO--,
wherein X may be, for example, F, Cl, and Br, and wherein R.sub.2
and R.sub.3 are as defined herein.
Inventors: |
Wu; Jinzi J.; (Dollard des
Ormeaux, CA) ; Stranix; Brent Richard;
(Pointe-Claire, CA) ; Ge; Michael; (St-Laurent,
CA) ; Milot; Guy; (Blainville, CA) ; Petrella;
Marco; (Montreal, CA) ; Panchal; Chandra;
(London, CA) |
Correspondence
Address: |
GODFREY & KAHN S.C.
780 NORTH WATER STREET
MILWAUKEE
WI
53202
US
|
Assignee: |
AMBRILIA BIOPHARMA INC.
Verdun
CA
H3E 1H4
|
Family ID: |
37214407 |
Appl. No.: |
11/380291 |
Filed: |
April 26, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60675082 |
Apr 27, 2005 |
|
|
|
Current U.S.
Class: |
514/237.5 ;
514/220; 514/249; 514/255.06; 514/263.34; 514/310; 514/313;
514/317; 514/357; 514/602 |
Current CPC
Class: |
A61K 31/661 20130101;
A61K 31/44 20130101; A61K 31/445 20130101; A61K 31/522 20130101;
A61K 31/135 20130101; A61K 31/551 20130101; A61K 31/513 20130101;
A61P 31/18 20180101; A61K 31/498 20130101; A61K 31/5375 20130101;
A61K 31/4196 20130101; A61K 31/47 20130101; A61K 45/06 20130101;
A61K 31/18 20130101; A61K 31/4965 20130101 |
Class at
Publication: |
514/237.5 ;
514/255.06; 514/313; 514/317; 514/357; 514/310; 514/249; 514/602;
514/263.34; 514/220 |
International
Class: |
A61K 31/5375 20060101
A61K031/5375; A61K 31/4965 20060101 A61K031/4965; A61K 31/498
20060101 A61K031/498; A61K 31/551 20060101 A61K031/551; A61K 31/445
20060101 A61K031/445; A61K 31/44 20060101 A61K031/44; A61K 31/522
20060101 A61K031/522; A61K 31/47 20060101 A61K031/47; A61K 31/18
20060101 A61K031/18 |
Claims
1. A pharmaceutical composition comprising: a) a compound of
formula I ##STR426## and pharmaceutically acceptable salts thereof,
wherein n is 3 or 4, wherein X and Y, the same or different, are
selected from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5,
--NHCOR.sub.4, --OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4,
and --CH.sub.2OH or X and Y together define an alkylenedioxy group
selected from the group consisting of a methylenedioxy group of
formula --OCH.sub.2O-- and an ethylenedioxy group of formula
--OCH.sub.2CH.sub.2O--, wherein R.sub.6 is selected from the group
consisting of a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof, wherein R.sub.3 is
selected from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, and a group of formula
R.sub.3A--CO--, R.sub.3A being selected from the group consisting
of a straight or branched alkyl group of 1 to 6 carbon atoms, a
cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof, an alkyloxy group of
1 to 6 carbon atoms, tetrahydro-3-furanyloxy, --CH.sub.2OH,
--CF.sub.3, --CH.sub.2CF.sub.3, --CH.sub.2CH.sub.2CF.sub.3,
pyrrolidinyl, piperidinyl, 4-morpholinyl, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 3-hydroxyphenyl, 4-hydroxyphenyl,
4-CH.sub.3OC.sub.6H.sub.4CH.sub.2--, CH.sub.3NH--,
(CH.sub.3).sub.2N--, (CH.sub.3CH.sub.2).sub.2N--,
(CH.sub.3CH.sub.2CH.sub.2).sub.2N--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--,
C.sub.6H.sub.5CH.sub.2O--, 2-pyrrolyl, 2-pyridyl, 3-pyridyl,
4-pyridyl-, 2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl,
1-isoquinolyl, 3-isoquinolyl, 2-quinoxalinyl, a phenyl group of
formula ##STR427## a picolyl group selected from the group
consisting of ##STR428## a picolyloxy group selected from the group
consisting of ##STR429## a substituted pyridyl group selected from
the group consisting of ##STR430## a group of formula ##STR431##
wherein X' and Y', the same or different, are selected from the
group consisting of H, a straight alkyl group of 1 to 6 carbon
atoms, a branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl
group of 3 to 6 carbon atoms, F, Cl, Br, l, --CF.sub.3, --NO.sub.2,
--NR.sub.4R.sub.5, --NHCOR.sub.4, --OR.sub.4, --SR.sub.4,
--COOR.sub.4, --COR.sub.4 and --CH.sub.2OH, wherein R.sub.4 and
R.sub.5, the same or different, are selected from the group
consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, and a cycloalkyl group
of 3 to 6 carbon atoms, wherein R.sub.2 is selected from the group
consisting of a diphenylmethyl group of formula IV ##STR432## a
naphthyl-1-CH.sub.2-- group of formula V ##STR433## a
naphthyl-2-CH.sub.2-- group of formula VI ##STR434## a
biphenylmethyl group of formula VII ##STR435## and an
anthryl-9-CH.sub.2-- group of formula VIII ##STR436## wherein
R.sub.1 is H or a physiologically cleavable unit and whereby upon
physiological conditions said compound is converted into an active
protease inhibitor; b) a cytochrome P450 monooxigenase inhibitor,
and; c) a pharmaceutically acceptable carrier.
2. The pharmaceutical composition of claim 1, wherein R.sub.1 is
selected from the group consisting of H, (HO).sub.2P(O) and
(MO).sub.2P(O), and a group of formula R.sub.1A--CO--, R.sub.1A
being selected from the group consisting of a straight or branched
alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to
6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms
in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl
part thereof, an alkyloxy group of 1 to 6 carbon atom,
--CH.sub.2OH, CH.sub.3O.sub.2C--, CH.sub.3O.sub.2CCH.sub.2--,
Acetyl-OCH.sub.2CH.sub.2--, HO.sub.2CCH2,2-hydroxyphenyl,
3-hydroxyphenyl, 4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula ##STR437##
a picolyl group selected from the group consisting of ##STR438## a
picolyloxy group selected from the group consisting of ##STR439## a
substituted pyridyl group selected from the group consisting of
##STR440## and a group of formula, ##STR441## wherein M is an
alkali metal or alkaline earth metal and wherein X', Y', R.sub.4
and R.sub.5 are as defined in claim 1.
3. A pharmaceutical composition comprising; a) a compound of
formula II, ##STR442## and pharmaceutically acceptable salts
thereof, wherein n is 3 or 4, wherein X and Y, the same or
different, are selected from the group consisting of H, a straight
alkyl group of 1 to 6 carbon atoms, a branched alkyl group of 3 to
6 carbon atoms, a cycloalkyl group of 3 to 6 carbon atoms, F, Cl,
Br, I, --CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2,
--NR.sub.4R.sub.5, --NHCOR.sub.4, --OR.sub.4, --SR.sub.4,
--COOR.sub.4, --COR.sub.4, and --CH.sub.2OH or X and Y together
define an alkylenedioxy group selected from the group consisting of
a methylenedioxy group of formula --OCH.sub.2O-- and an
ethylenedioxy group of formula --OCH.sub.2CH.sub.2O--, wherein
R.sub.6 is selected from the group consisting of a straight alkyl
group of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6
carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in
the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl
part thereof, wherein R.sub.3 is selected from the group consisting
of H, a straight alkyl group of 1 to 6 carbon atoms, a branched
alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of 3 to 6
carbon atoms, and a group of formula R.sub.3A--CO--, R.sub.3A being
selected from the group consisting of a straight or branched alkyl
group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6
carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in
the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl
part thereof, an alkyloxy group of 1 to 6 carbon atoms,
tetrahydro-3-furanyloxy, --CH.sub.2OH, --CF.sub.3,
--CH.sub.2CF.sub.3, --CH.sub.2CH.sub.2CF.sub.3, pyrrolidinyl,
piperidinyl, 4-morpholinyl, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 3-hydroxyphenyl, 4-hydroxyphenyl,
4-CH.sub.3OC.sub.6H.sub.4CH.sub.2--, CH.sub.3NH--,
(CH.sub.3).sub.2N--, (CH.sub.3CH.sub.2).sub.2N--,
(CH.sub.3CH.sub.2CH.sub.2).sub.2N--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--,
C.sub.6H.sub.5CH.sub.2O--, 2-pyrrolyl, 2-pyridyl, 3-pyridyl,
4-pyridyl-, 2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl,
1-isoquinolyl, 3-isoquinolyl, 2-quinoxalinyl, a phenyl group of
formula ##STR443## a picolyl group selected from the group
consisting of ##STR444## a picolyloxy group selected from the group
consisting of ##STR445## a substituted pyridyl group selected from
the group consisting of ##STR446## a group of formula ##STR447##
wherein X' and Y', the same or different, are selected from the
group consisting of H, a straight alkyl group of 1 to 6 carbon
atoms, a branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl
group of 3 to 6 carbon atoms, F, Cl, Br, I, --CF.sub.3, --NO.sub.2,
--NR.sub.4R.sub.5, --NHCOR.sub.4, --OR.sub.4, --SR.sub.4,
--COOR.sub.4, --COR.sub.4 and --CH.sub.2OH, wherein R.sub.4 and
R.sub.5, the same or different, are selected from the group
consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, and a cycloalkyl group
of 3 to 6 carbon atoms, wherein R.sub.2 is selected from the group
consisting of a diphenylmethyl group of formula IV ##STR448## a
naphthyl-1-CH.sub.2-- group of formula V ##STR449## a
naphthyl-2-CH.sub.2-- group of formula VI ##STR450## a
biphenylmethyl group of formula VII ##STR451## and an
anthryl-9-CH.sub.2-- group of formula VIII ##STR452## wherein
R.sub.1 is H or a physiologically cleavable unit, whereby upon
physiological conditions said compound is converted into an active
protease inhibitor; b) a cytochrome P450 monooxigenase inhibitor,
and; c) a pharmaceutically acceptable carrier.
4. The pharmaceutical composition of claim 3, wherein R.sub.1 is
selected from the group consisting of H, (HO).sub.2P(O) and
(MO).sub.2P(O), and a group of formula R.sub.1A--CO--, R.sub.1A
being selected from the group consisting of a straight or branched
alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to
6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms
in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl
part thereof, an alkyloxy group of 1 to 6 carbon atoms,
--CH.sub.2OH, CH.sub.3O.sub.2C--, CH.sub.3O.sub.2CCH.sub.2--,
Acetyl-OCH.sub.2CH.sub.2--, HO.sub.2CCH.sub.2--, 2-hydroxyphenyl,
3-hydroxyphenyl, 4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula ##STR453##
a picolyl group selected from the group consisting of ##STR454## a
picolyloxy group selected from the group consisting of ##STR455## a
substituted pyridyl group selected from the group consisting of
##STR456## and a group of formula, ##STR457## wherein M is an
alkali metal or alkaline earth metal and wherein X', Y', R.sub.4
and R.sub.5 are as defined in claim 1.
5. The pharmaceutical composition of claim 4, wherein R.sub.6 is
iso-butyl and n is 3.
6. The pharmaceutical composition of claim 4, wherein R.sub.6 is
isobutyl and n is 4.
7. The pharmaceutical composition of claim 6, wherein R.sub.1 is
selected from the group consisting of H, (HO).sub.2P(O) and
(NaO).sub.2P(O).
8. The pharmaceutical composition of claim 6, wherein R.sub.1 is
selected from the group consisting of CH.sub.3CO, 3-pyridyl-CO,
(CH.sub.3).sub.2NCH.sub.2CO and
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
9. The pharmaceutical composition of claim 7, wherein R.sub.3 is
selected from the group consisting of CH.sub.3CO, CH.sub.3O--CO,
(CH.sub.3).sub.2N--CO, 3-pyridyl-CO, 4-pyridyl-CO and
4-morpholine-Co.
10. The pharmaceutical composition of claim 8, wherein R.sub.3 is
selected from the group consisting of CH.sub.3CO, CH.sub.3O--CO,
(CH.sub.3).sub.2N--CO, 3-pyridyl-CO, 4-pyridyl-CO and
4-morpholine-Co.
11. The pharmaceutical composition of claim 9, wherein X is
4-NH.sub.2 and Y is H or F.
12. The pharmaceutical composition of claim 10, wherein X is
4-NH.sub.2 and Y is H or F.
13. The pharmaceutical composition of claim 11, wherein R.sub.2 is
selected from the group consisting of a diphenylmethyl group of
formula IV, a naphthyl-1-CH.sub.2-- group of formula V, a
naphthyl-2-CH.sub.2-- group of formula VI, a biphenylmethyl group
of formula VII and an anthryl-9-CH.sub.2-- group of formula
VIII.
14. The pharmaceutical composition of claim 12, wherein R.sub.2 is
selected from the group consisting of a diphenylmethyl group of
formula IV, a naphthyl-1-CH.sub.2-- group of formula V, a
naphthyl-2-CH.sub.2-- group of formula VI, a biphenylmethyl group
of formula VII and an anthryl-9-CH.sub.2-- group of formula
VIII.
15. The pharmaceutical composition of claim 13, wherein R.sub.2 is
selected from the group consisting of a diphenylmethyl group of
formula IV, a naphthyl-1-CH.sub.2-- group of formula V, and a
naphthyl-2-CH.sub.2-- group of formula VI.
16. The pharmaceutical composition of claim 14, wherein R.sub.2 is
selected from the group consisting of a diphenylmethyl group of
formula IV, a naphthyl-1-CH.sub.2-- group of formula V, and a
naphthyl-2-CH.sub.2-- group of formula VI.
17. The pharmaceutical composition of claim 15, wherein X' and Y'
is H.
18. The pharmaceutical composition of claim 16, wherein X' and Y'
is H.
19. The pharmaceutical composition of claim 6 wherein R.sub.2 is a
diphenylmethyl group of formula IV.
20. The pharmaceutical composition of claim 19, wherein R.sub.1 is
selected from the group consisting of H, (HO).sub.2P(O) and
(NaO).sub.2P(O).
21. The pharmaceutical composition of claim 19, wherein R.sub.1 is
selected from the group consisting of CH.sub.3CO, 3-pyridyl-CO,
(CH.sub.3).sub.2NCH.sub.2CO and
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
22. The pharmaceutical composition of claim 20, wherein R.sub.3 is
selected from the group consisting of CH.sub.3CO, CH.sub.3O--CO,
(CH.sub.3).sub.2N--CO, 3-pyridyl-CO, 4-pyridyl-CO and
4-morpholine-CO.
23. The pharmaceutical composition of claim 21, wherein R.sub.3 is
selected from the group consisting of CH.sub.3CO, CH.sub.3O--CO,
(CH.sub.3).sub.2N--CO, 3-pyridyl-CO, 4-pyridyl-CO and
4-morpholine-CO.
24. The pharmaceutical composition of claim 22, wherein X is
4-NH.sub.2 and Y is H or F.
25. The pharmaceutical composition of claim 23, wherein X is
4-NH.sub.2 and Y is H or F.
26. The pharmaceutical composition of claim 22, wherein X is
4-NH.sub.2, Y is H, X' is H, Y' is H and R.sub.3 is
CH.sub.3O--CO.
27. The pharmaceutical composition of claim 26, wherein R.sub.1 is
(HO).sub.2P(O).
28. The pharmaceutical composition of claim 26, wherein R.sub.1 is
(NaO).sub.2P(O).
29. The pharmaceutical composition of claim 26, wherein R.sub.1 is
H.
30. The pharmaceutical composition of claim 22, wherein X is
4-NH.sub.2, Y is 3-F, X' is H, Y' is H and R.sub.3 is
CH.sub.3O--CO.
31. The pharmaceutical composition of claim 30, wherein R.sub.1 is
(HO).sub.2P(O).
32. The pharmaceutical composition of claim 30, wherein R.sub.1 is
(NaO).sub.2P(O).
33. The pharmaceutical composition of claim 30, wherein R.sub.1 is
H.
34. The pharmaceutical composition of claim 22, wherein X is
4-NH.sub.2, Y is H or 3-F, X' is H, Y' is H and R.sub.3 is
CH.sub.3CO.
35. The pharmaceutical composition of claim 34, wherein R.sub.1 is
(HO).sub.2P(O).
36. The pharmaceutical composition of claim 34, wherein R.sub.1 is
(NaO).sub.2P(O).
37. The pharmaceutical composition of claim 34, wherein R.sub.1 is
H.
38. The pharmaceutical composition of claim 22, wherein X is
4-NH.sub.2, Y is H or 3-F, X' is H, Y' is H and R.sub.3 is
4-morpholine-CO.
39. The pharmaceutical composition of claim 23, wherein X is
4-NH.sub.2, Y is H, X' is H, Y' is H and R.sub.3 is
CH.sub.3O--CO.
40. The pharmaceutical composition of claim 39, wherein R.sub.1 is
3-pyridyl-CO.
41. The pharmaceutical composition of claim 39, wherein R.sub.1 is
(CH.sub.3).sub.2NCH.sub.2CO.
42. The pharmaceutical composition of claim 39, wherein R.sub.1 is
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
43. The pharmaceutical composition of claim 39, wherein R.sub.1 is
CH.sub.3CO.
44. The pharmaceutical composition of claim 17, wherein R.sub.2 is
Naphtyl-1-CH.sub.2--, Y is H and R.sub.3 is 4-morpholine-CO.
45. The pharmaceutical composition of claim 17, wherein R.sub.2 is
Naphtyl-2-CH.sub.2--, Y is H and R.sub.3 is CH.sub.3O--CO.
46. The pharmaceutical composition of claim 1, wherein said
cytochrome P450 monooxigenase inhibitor is selected from the group
consisting of ritonavir (RTV), ketoconazole, fluconazole,
nefazodone, fluvoxamine, fluoxetine, macrolide antibiotics,
sertraline sulfaphenazole and erythromycin.
47. The pharmaceutical composition of claim 1, wherein said
composition is administered orally.
48. The pharmaceutical composition of claim 1, wherein said
composition is administered twice-daily.
49. A kit for treating or preventing an HIV infection or for
treating or preventing AIDS, the kit comprising a) a first
container containing a compound of formula I; ##STR458## or a
pharmaceutically acceptable salts thereof wherein n, X, Y, X', Y',
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5 and R.sub.6 are as
defined in claim 1 and a second container containing a cytochrome
P450 monooxigenase inhibitor or; b) a container comprising both the
compound of formula I and the CYP450 inhibitor.
50. The kit as defined in claim 49, wherein R.sub.1 is as defined
in claim 2.
51. The kit as defined in any claim 49, wherein said cytochrome
P450 monooxigenase inhibitor is selected from the group consisting
of ritonavir (RTV), ketoconazole, fluconazole, nefazodone,
fluvoxamine, fluoxetine, macrolide antibiotics, sertraline
sulfaphenazole and erythromycin.
52. A method of treating or preventing an HIV infection or of
treating or preventing AIDS, the method comprising administering a
pharmaceutical composition as defined in claim 1 to a mammal in
need thereof.
53. The method of claim 52, wherein said composition is
administered orally.
54. The method of claim 52, wherein said composition is
administered twice-daily.
55. The use of a pharmaceutical composition as defined in claim 1
for the treatment or prevention of an HIV infection or for the
treatment or prevention of AIDS.
56. A method of treating or preventing an HIV infection or of
treating or preventing AIDS, the method comprising administering a)
a compound of formula I ##STR459## wherein n, X, Y, X', Y',
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5 and R.sub.6 are as
defined in claim 1 or a pharmaceutically acceptable salts thereof,
and; b) one or more CYP450 inhibitor in an amount which is
sufficient to reduce the metabolism of the compound of formula
I.
57. The method as defined in claim 56, wherein said CYP450
inhibitor is selected from the group consisting of ritonavir (RTV),
ketoconazole, fluconazole, nefazodone, fluvoxamine, fluoxetine,
macrolide antibiotics, sertraline sulfaphenazole and
erythromycin.
58. The method of claim 56, wherein administration of the compound
of formula I and the CYP450 inhibitor is performed separately,
simultaneously or sequentially.
59. The method of claim 58, wherein the administration of the
compound of formula I and the CYP450 inhibitor is performed by
administering a) a first pharmaceutical composition comprising a
compound of formula I and a pharmaceutically acceptable carrier and
b) a second pharmaceutical composition comprising a CYP450
inhibitor and a pharmaceutically acceptable carrier.
60. The method of claim 58, wherein the administration of the
compound of formula I and the CYP450 inhibitor is performed by
administering a single pharmaceutical composition comprising a
compound of formula I, a CYP450 inhibitor and a pharmaceutically
acceptable carrier.
61. The method of claim 60, wherein said CYP450 inhibitor is
ritonavir.
62. A method for improving the pharmacokinetics of a compound of
formula I as defined in claim 1, the method comprising
administering to a human in need thereof, the compound of formula I
and an amount of a CYP450 inhibitor effective to inhibit cytochrome
P450 monooxygenase.
63. The pharmaceutical composition of claim 1, wherein the ratio of
the compound of formula I over the cytochrome P450 monooxygenase
inhibitor is between about 1:1 to about 10:1.
64. The pharmaceutical composition of claim 63, wherein the ratio
is between about 3:1 and about 6:1.
65. A method of inhibiting an HIV having a reduced susceptibility
to a protease inhibitor other than the protease inhibitor defined
by formula I, the method comprising administering a compound of
formula I alone or in combination with a CYP450 inhibitor to an
individual in need thereof.
66. The method of claim 65, wherein said HIV is HIV-1.
67. The method of claim 66, wherein said HIV has a reduced
susceptibility to one or more of a protease inhibitor selected from
the group consisting of Atazanavir, Amprenavir, Indinavir,
Lopinavir, Nelfinavir, Ritonavir and Saquinavir.
68. The method of claim 67, wherein said HIV-1 possesses an
aspartyl protease having one or more mutations.
Description
RELATED APPLICATION
[0001] The present application seeks priority from U.S. Provisional
application 60/675,082, which is incorporated by reference in its
entirety, as if fully set forth herein.
TECHNICAL FIELD OF THE INVENTION
[0002] This invention relates to method for improving the
pharmacokinetics of protease inhibitors and protease inhibitor
precursors and related pharmaceutical compositions. More
particularly, the present invention relates to method for improving
the pharmacokinetics of protease inhibitors and protease inhibitor
precursors by co-administering a cytochrome P450 monooxygenase
inhibitor.
BACKGROUND OF THE INVENTION
[0003] Inhibitors of the HIV viral protease have been developed
relatively recently and their use began only in 1996. Currently,
they are considered the most effective drugs against HIV infection.
Unfortunately, most current proteases inhibitors are relatively
large hydrophobic molecules that possess rather low
bioavailability. A high pill burden is therefore required to attain
the therapeutic dose in a patient. This is a deterrent, which too
often results in patient non-compliance and inadequate treatment
results. This situation leads to sub-optimal therapeutic drug
concentration that in turns leads to the development of HIV
resistant strains. Consequently, there is an urgent need to improve
the solubility and bioavailability of proteases inhibitors.
[0004] Examples of improved compounds have been developed in the
form of prodrugs of aspartyl protease inhibitors such as described,
for example, in U.S. Pat. No. 6,436,989 to Hale et al, the entire
content of which is incorporated herein by reference. This patent
shows a novel class of molecules characterized by favourable
aqueous solubility, high oral bioavailability and facile in vivo
generation of the active ingredient. However, it is well known that
HIV has the ability to develop resistance to the currently
available drugs. Thus, there is a need for alternative HIV protease
inhibitors active towards wild-type and resistant viral strains.
Thus, molecules derived from current HIV protease inhibitors
showing enhanced solubility and bioavailability is desirable to
fight resistant viral strains.
[0005] A unique class of aromatic derivatives which are inhibitors
of aspartyl proteases is described in U.S. Pat. No. 6,632,816 to
Stranix et al, the entire content of which is incorporated herein
by reference. This patent includes, more particularly, N-synthetic
amino acid substituted L-lysine derivatives possessing potent
aspartyl protease inhibitory properties. However, it would be
advantageous to improve these derivatives by enhancing aqueous
solubility and bioavailability in order to reduce the pill burden
and to favour patient's compliance. Since it is challenging to
generate active protease inhibitors, specifically toward wild-type
and resistant strains, the formation of derivatives of original HIV
protease inhibitors such as inhibitors described in U.S. Pat. No.
6,632,816 to Stranix et al, known to be active toward resistant
strains represents a viable route with considerable advantages.
More particularly, generation of compounds and formulations with
enhanced aqueous solubility, oral bioavailability, time of duration
and formulation properties along with other advantages is desirable
in the development of an effective drug. Protease inhibitors with
improved pharmacokinetics are therefore desirable.
SUMMARY OF THE INVENTION
[0006] Lysine-based compounds with increased solubility and
improved oral bioavailability are described herein and have been
described in U.S. patent application Ser. No. 10/902,935 filed on
Aug. 2, 2004 and published on Feb. 2, 2006 under No.
2006/0025592A1, the entire content of which is incorporated herein
by reference. These compounds may readily be cleaved in vivo to
release an active ingredient which has an affinity for aspartyl
proteases and which may act as a protease inhibitor. More
particularly, the active ingredient may bind, for example, to an
HIV aspartyl protease (U.S. Pat. No. 6,632,816) and may inhibit
this enzyme. The Lysine-based compounds are also referred herein as
a protease inhibitor precursor.
[0007] Upon in vivo physiological conditions (e.g., metabolic,
enteric and/or gastrointestinal conditions, etc.) the Lysine-based
compounds, allow for the release of a protease inhibitor (e.g.,
aspartyl protease inhibitor). The Lysine-based compounds may thus
serve as means for improving the solubility and/or bioavailability
of protease inhibitors and therefore may reduce the pill burden
and/or reduce dosages needed for inhibition. Improved treatment of
HIV-infected patients and favourable patient's compliance may
consequently occur.
[0008] The Lysine-based compounds described herein may be used
alone or in combination with other therapeutic or prophylactic
agents for the treatment or prophylaxis of HIV infection.
[0009] The active ingredient when released from the Lysine-based
compound may act, for example, on aspartyl protease of HIV-1
including mutated and non-mutated HIV-1 viral strain (e.g., NL4.3)
as well as on protease of HIV-2 (mutated or non-mutated) or even on
protease of related viruses (e.g. retroviruses (e.g. SIV, etc.)
etc.).
[0010] Other means to increase the pharmacokinetics of Lysine-based
compounds described herein or of the active ingredients disclosed
in U.S. Pat. No. 6,632,816 to Stranix et al, are investigated
herein.
[0011] In accordance with the present invention, there is provided
a method of improving the pharmacokinetics of a Lysine-based
compound or of the active ingredients disclosed in U.S. Pat. No.
6,632,816 to Stranix et al. The method may comprise administering,
to an individual in need thereof, a cytochrome P450 monooxygenase
inhibitor (or a pharmaceutically acceptable salt thereof) in
combination with a Lysine-based compound (or a pharmaceutically
acceptable salt thereof) and/or with an active ingredient as
disclosed in U.S. Pat. No. 6,632,816 (or a pharmaceutically
acceptable salt thereof).
[0012] The method may comprise administering the cytochrome P450
monooxygenase inhibitor and the protease inhibitor or protease
inhibitor precursor either separately, simultaneously or
sequentially. The method may also comprise administering the
cytochrome P450 monooxygenase inhibitor and the protease inhibitor
or protease inhibitor precursor at different time interval.
[0013] For example, when administering a combination of a
Lysine-based compound (or a pharmaceutically acceptable salt
thereof) and a cytochrome P450 monooxygenase inhibitor (or a
pharmaceutically acceptable salt thereof) the two therapeutic
agents may be formulated as separate composition which may be
administered separately at the same time or at different times,
using the same route of administration or different routes of
administration, at the same administration site or at different
administration sites etc. Alternatively, the therapeutic agents may
be administered as a single composition either orally, by
injection, etc.
[0014] In accordance with the present invention, the CYP450
inhibitor may be a CYP450-3A inhibitor.
[0015] Cytochrome P450 monooxygenase inhibitor (CYP450 inhibitor)
which may be used to increase the pharmacokinetics of Lysine-based
compounds or of the active ingredients disclosed in U.S. Pat. No.
6,632,816 to Stranix et al., include for example, ritonavir (RTV),
ketoconazole, fluconazole, nefazodone, fluvoxamine, fluoxetine,
macrolide antibiotics, sertraline sulfaphenazole, erythromycin
etc.
[0016] The present invention further provides a method for
treating, preventing, reducing the risk or probability of HIV
infection or reducing HIV burden. The invention also provides a
method for treating, preventing or reducing the risk or probability
of developing AIDS, for delaying the apparition of AIDS or reducing
AIDS symptoms. The method may comprise administering a Lysine-based
compound (or a pharmaceutically acceptable salt thereof) as
described herein or the active ingredients disclosed in U.S. Pat.
No. 6,632,816 to Stranix et al, and a CYP450 inhibitor (or a
pharmaceutically acceptable salt thereof).
[0017] In yet a further aspect, the present invention provides a
method of treating, preventing or reducing the risk or probability
of HIV infection or of reducing HIV burden. The invention also
provides a method for treating, preventing or reducing the risk or
probability of developing acquired immunodeficiency syndrome
(AIDS), for delaying the apparition of AIDS, or reducing AIDS
symptoms. The method may comprise administering at least one
compound of formula I, II, IIa, IIb, IIc, IIA, IIA' or
pharmaceutically acceptable salts or derivatives thereof or the
active ingredients disclosed in U.S. Pat. No. 6,632,816 to Stranix
et al, pharmaceutically acceptable salts or derivatives thereof, or
mixture of any of these compounds to a mammal in need thereof in
combination with a CYP450 inhibitor.
[0018] The present invention also relates in a further aspect
thereof to a pharmaceutical composition comprising a) a
Lysine-based compound (or a pharmaceutically acceptable salt or
derivatives thereof) as described herein or an active ingredient as
disclosed in U.S. Pat. No. 6,632,816 to Stranix et al. (or
pharmaceutically acceptable salts or derivatives thereof or mixture
of any one of these compounds thereof; b) a CYP450 inhibitor (or a
pharmaceutically acceptable salt thereof; and c) a pharmaceutically
acceptable carrier.
[0019] In yet another aspect, the present invention relates to a
pharmaceutical composition comprising at least one compound of
formula I, II, IIa, IIb, IIc, IIA, IIA' as described herein (or
pharmaceutically acceptable salts or derivatives thereof or an
active ingredient as disclosed in U.S. Pat. No. 6,632,816 to
Stranix et al., (or pharmaceutically acceptable salts or
derivatives thereof or mixture of any of these compounds and a
CYP450 inhibitor (or a pharmaceutically acceptable salt thereof.
The pharmaceutical composition may also comprise a pharmaceutically
acceptable carrier. The pharmaceutical composition may comprise,
for example, a pharmaceutically effective amount of such one or
more compounds or as applicable, pharmaceutically acceptable
ammonium salts thereof.
[0020] More particularly, the pharmaceutical composition may
consist essentially of at least one compound of formula I, II, IIa,
IIb, IIc, IIA, IIA' as described herein or pharmaceutically
acceptable salts or derivatives thereof or the active ingredients
disclosed in U.S. Pat. No. 6,632,816 to Stranix et al, or mixture
of any of these compounds and a CYP450 inhibitor (or a
pharmaceutically acceptable salt thereof).
[0021] Additionally, the pharmaceutical composition may consist of
at least one compound of formula I, II, IIa, IIb, IIc, IIA, IIA' as
described herein or pharmaceutically acceptable salts or
derivatives thereof or the active ingredients disclosed in U.S.
Pat. No. 6,632,816 to Stranix et al, or mixture of any of these
compounds and a CYP450 inhibitor (or a pharmaceutically acceptable
salt thereof.
[0022] The present invention also relates in an additional aspect
thereof to the use of a combination of a) a Lysine-based compound
(or a pharmaceutically acceptable salt thereof) as described herein
or the active ingredients disclosed in U.S. Pat. No. 6,632,816 to
Stranix et al, (or a pharmaceutically acceptable salt thereof) or
mixture of any of these compounds and b) a CYP450 inhibitor (or a
pharmaceutically acceptable salt thereof) in the making of a
pharmaceutical composition for the treatment or prevention of a
retroviral infection (e.g., HTLV, HIV, i.e., HIV-1, HIV-2) or for
the treatment or prevention of acquired immunodeficiency syndrome
(AIDS).
[0023] In an additional aspect, the present invention relates to
the use of at least one compound of formula I, II, IIa, IIb, IIc,
IIA, IIA' as described herein or pharmaceutically acceptable salts
or derivatives thereof or the active ingredients disclosed in U.S.
Pat. No. 6,632,816 to Stranix et al, or mixture of any of these
compounds and a CYP450 inhibitor (or a pharmaceutically acceptable
salt thereof) in the manufacture of a drug (or pharmaceutical
composition) for the treatment or prevention of an HIV infection
(for reducing the risk or probability of HIV infection), or again
for reducing HIV burden in a mammal in need thereof or for the
treatment or prevention of acquired immunodeficiency syndrome
(AIDS) (for reducing the risk or probability of developing AIDS)
for delaying the apparition of AIDS (symptoms) or for reducing AIDS
symptoms in a mammal in need thereof.
[0024] A "mammal in need" is to be understood herein, without
limitation, as an individual infected with HIV (i.e., at any stage
of HIV infection, e.g., primary infection, symptomatic,
asymptomatic, AIDS) or at risk of having an HIV infection. A
"mammal in need" therefore, may comprise, in addition to
individuals who has an acute HIV infection, a chronic HIV infection
or AIDS, individuals (e.g., health care worker, an individual who
had unprotected sexual intercourse, policemen, etc.) who may have,
for example, come into contact with a possible source of
contamination with HIV.
[0025] The present invention, additionally provides the use of a
combination of a Lysine-based compound (or a pharmaceutically
acceptable salt thereof) as described herein or the use of an
active ingredient disclosed in U.S. Pat. No. 6,632,816 to Stranix
et al, (or a pharmaceutically acceptable salt thereof) and a CYP450
inhibitor (or a pharmaceutically acceptable salt thereof) for the
treatment, prevention or for reducing the risk or probability of a
retroviral infection in a mammal in need thereof.
[0026] More particularly, the present invention relates to the use
of a combination of at least one compound of formula I, II, IIa,
IIb, IIc, IIA, IIA' as described herein or pharmaceutically
acceptable salts or derivatives thereof or the active ingredients
disclosed in U.S. Pat. No. 6,632,816 to Stranix et al, (or a
pharmaceutically acceptable salt thereof) or mixture of any of
these compounds and a CYP450 inhibitor in the treatment, prevention
or for reducing the risk or probability of an HIV infection in a
mammal in need thereof or for the treatment, prevention or for
reducing the risk or probability of developing acquired
immunodeficiency syndrome (AIDS).
[0027] Exemplary embodiments of Lysine-based compounds encompassed
by the present invention and which may be used in combination with
a CYP450 inhibitor for the methods, pharmaceutical compositions,
kits and uses described herein, may include, for example, a
compound of formula I: ##STR2##
[0028] pharmaceutically acceptable salts and derivatives thereof
(e.g., for example, when the compound of the present invention
comprises an amino group, the pharmaceutically acceptable salt may
be an ammonium salt),
[0029] wherein n may be, for example, 3 or 4,
[0030] wherein X and Y, the same or different, may be selected, for
example, from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5,
--NHCOR.sub.4, --OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4,
and --CH.sub.2OH or X and Y together define an alkylenedioxy group
selected from the group consisting of a methylenedioxy group of
formula --OCH.sub.2O-- and an ethylenedioxy group of formula
--OCH.sub.2CH.sub.2O--,
[0031] wherein R.sub.6 may be selected, for example, from the group
consisting of a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof,
[0032] wherein R.sub.3 may be selected, for example, from the group
consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of
3 to 6 carbon atoms, and a group of formula R.sub.3A--CO--, wherein
R.sub.3A may be selected, for example, from the group consisting of
a straight or branched alkyl group of 1 to 6 carbon atoms (e.g.
methyl, ethyl-, propyl, iso-propyl, butyl, iso-butyl, tert-butyl,
tert-butyl-CH.sub.2--, etc.), a cycloalkyl group having 3 to 6
carbon atoms (e.g. cyclopropyl-, cyclohexyl-etc.), a
cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl
part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
(e.g. cyclopropyl-CH.sub.2--, cyclohexyl-CH.sub.2--, etc.), an
alkyloxy group of 1 to 6 carbon atoms (e.g. CH.sub.3O--,
CH.sub.3CH.sub.2O--, iso-butylO-, tert-butylO-(Boc), etc.),
tetrahydro-3-furanyloxy, --CH.sub.2OH, --CF.sub.3,
--CH.sub.2CF.sub.3, --CH.sub.2CH.sub.2CF.sub.3, pyrrolidinyl,
piperidinyl, 4-morpholinyl, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 3-hydroxyphenyl, 4-hydroxyphenyl,
4-CH.sub.3OC.sub.6H.sub.4CH.sub.2--, CH.sub.3NH--,
(CH.sub.3).sub.2N--, (CH.sub.3CH.sub.2).sub.2N--,
(CH.sub.3CH.sub.2CH.sub.2).sub.2N--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--,
C.sub.6H.sub.5CH.sub.2O--, 2-pyrrolyl, 2-pyridyl, 3-pyridyl,
4-pyridyl-, 2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl,
1-isoquinolyl, 3-isoquinolyl, 2-quinoxalinyl, a phenyl group of
formula ##STR3##
[0033] a picolyl group selected from the group consisting of
##STR4##
[0034] a picolyloxy group selected from the group consisting of
##STR5##
[0035] a substituted pyridyl group selected from the group
consisting of ##STR6##
[0036] and a group of formula, ##STR7##
[0037] wherein X' and Y', the same or different, may be selected,
for example, from the group consisting of H, a straight alkyl group
of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon
atoms, a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4 and
--CH.sub.2OH,
[0038] wherein R.sub.4 and R.sub.5, the same or different, may be
selected, for example, from the group consisting of H, a straight
alkyl group of 1 to 6 carbon atoms, a branched alkyl group of 3 to
6 carbon atoms, and a cycloalkyl group of 3 to 6 carbon atoms,
[0039] wherein R.sub.2 may be selected, for example, from the group
consisting of a diphenylmethyl group of formula IV ##STR8##
[0040] a naphthyl-1-CH.sub.2-- group of formula V ##STR9##
[0041] a naphthyl-2-CH.sub.2-- group of formula VI ##STR10##
[0042] a biphenylmethyl group of formula VI ##STR11##
[0043] and an anthryl-9-CH.sub.2-- group of formula VIII
##STR12##
[0044] and wherein R.sub.1 may be a cleavable unit (e.g., a
physiologically cleavable unit), whereby upon cleavage of the unit,
the compound releases a protease inhibitor (an HIV protease
inhibitor), provided that R.sub.1 is not H. For example, R.sub.1
may be an enzymatically or metabolically cleavable unit or
hydrolysable bond which may be cleaved under enteric and/or
gastrointestinal conditions (pH) or other physiological
conditions.
[0045] In accordance with the present invention, R.sub.1 may be
selected, for example, from the group consisting of (HO).sub.2P(O)
and (MO).sub.2P(O), wherein M is an alkali metal (e.g. Na, K, Cs,
etc) or alkaline earth metal (Ca, Mg, etc.).
[0046] Further in accordance with the present invention, R.sub.1
may be a group of formula R.sub.1A--CO--, wherein R.sub.1A may be
selected, for example, from the group consisting of a straight or
branched alkyl group of 1 to 6 carbon atoms (e.g. methyl, ethyl,
propyl, iso-propyl, butyl, iso-butyl, tert-butyl,
tert-butyl-CH.sub.2--, etc.), a cycloalkyl group having 3 to 6
carbon atoms (e.g. cyclopropyl-, cyclohexyl- etc.), a
cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl
part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
(e.g. cyclopropyl-CH.sub.2--, cyclohexyl-CH.sub.2--, etc.), an
alkyloxy group of 1 to 6 carbon atoms (e.g. CH.sub.3O--,
CH.sub.3CH.sub.2O--, iso-butylO-, tert-butylO--(Boc), etc.),
--CH.sub.2OH, CH.sub.3O.sub.2C--, CH.sub.3O.sub.2CCH.sub.2--,
Acetyl-OCH.sub.2CH.sub.2--, HO.sub.2CCH.sub.2--, 2-hydroxyphenyl,
3-hydroxyphenyl, 4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula
##STR13##
[0047] a picolyl group selected from the group consisting of
##STR14##
[0048] a picolyloxy group selected from the group consisting of
##STR15##
[0049] a substituted pyridyl group selected from the group
consisting of ##STR16##
[0050] and a group of formula, ##STR17##
[0051] wherein X', Y', R.sub.4 and R.sub.5 are as defined
herein.
[0052] Alternatively, active ingredients disclosed in U.S. Pat. No.
6,632,816 to Stranix et al, which may be used in combination with a
CYP450 inhibitor in the methods, pharmaceutical compositions, kits
and uses described herein, are also defined by formula I, II, IIa,
IIb, IIc, IIA or IIA', (or a salt thereof), wherein R.sub.1 is H
and R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, n, X, Y, X', Y'
are as defined herein.
[0053] More particularly, the present invention relates to a
pharmaceutical composition which may comprise:
[0054] a) a compound of formula I as described herein or a
pharmaceutically acceptable salt thereof,
[0055] b) a cytochrome P450 monooxigenase inhibitor, and;
[0056] c) a pharmaceutically acceptable carrier;
[0057] where the compound of formula I is represented by;
##STR18##
[0058] wherein n may be 3 or 4,
[0059] wherein X and Y, the same or different, may be selected, for
example, from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5,
--NHCOR.sub.4, --OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4,
and --CH.sub.2OH or X and Y together may define an alkylenedioxy
group which may be selected, for example, from the group consisting
of a methylenedioxy group of formula --OCH.sub.2O-- and an
ethylenedioxy group of formula --OCH.sub.2CH.sub.2O--,
[0060] wherein R.sub.6 may be selected from the group consisting of
a straight alkyl group of 1 to 6 carbon atoms, a branched alkyl
group of 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6
carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms
in the alkyl part thereof,
[0061] wherein R.sub.3 may be selected, for example, from the group
consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of
3 to 6 carbon atoms, and a group of formula R.sub.3A--CO--, where
R.sub.3A may be selected from the group consisting of a straight or
branched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group
having 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6
carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms
in the alkyl part thereof, an alkyloxy group of 1 to 6 carbon
atoms, tetrahydro-3-furanyloxy, --CH.sub.2OH, --CF.sub.3,
--CH.sub.2CF.sub.3, --CH.sub.2CH.sub.2CF.sub.3, pyrrolidinyl,
piperidinyl, 4-morpholinyl, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 3-hydroxyphenyl, 4-hydroxyphenyl,
4-CH.sub.3OC.sub.6H.sub.4CH.sub.2--, CH.sub.3NH--,
(CH.sub.3).sub.2N--, (CH.sub.3CH.sub.2).sub.2N--,
(CH.sub.3CH.sub.2CH.sub.2).sub.2N--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--,
C.sub.6H.sub.5CH.sub.2O--, 2-pyrrolyl, 2-pyridyl, 3-pyridyl,
4-pyridyl-, 2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl,
1-isoquinolyl, 3-isoquinolyl, 2-quinoxalinyl, a phenyl group of
formula ##STR19##
[0062] a picolyl group which may be selected from the group
consisting of ##STR20##
[0063] a picolyloxy group which may be selected from the group
consisting of ##STR21##
[0064] a substituted pyridyl group which may be selected from the
group consisting of ##STR22##
[0065] a group of formula ##STR23##
[0066] wherein X' and Y', the same or different, may be selected,
for example, from the group consisting of H, a straight alkyl group
of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon
atoms, a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4 and
--CH.sub.2OH,
[0067] wherein R.sub.4 and R.sub.5, the same or different, may be
selected from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
and a cycloalkyl group of 3 to 6 carbon atoms,
[0068] wherein R.sub.2 may be selected from the group consisting of
a diphenylmethyl group of formula IV ##STR24##
[0069] a naphthyl-1-CH.sub.2-- group of formula V ##STR25##
[0070] a naphthyl-2-CH.sub.2-- group of formula VI ##STR26##
[0071] a biphenylmethyl group of formula VII ##STR27##
[0072] and an anthryl-9-CH.sub.2-- group of formula VIII
##STR28##
[0073] wherein R.sub.1 may be H or a physiologically cleavable
unit, whereby upon (in vivo) physiological conditions the compound
may be converted into an active protease inhibitor. For example,
upon cleavage of the physiologically cleavable unit, the compound
may be able to release a protease inhibitor.
[0074] In accordance with the present invention, R.sub.1 may be
selected, for example, from the group consisting of H,
(HO).sub.2P(O) and (MO).sub.2P(O) (wherein M may be, for example,
an alkali metal or alkaline earth metal) and a group of formula
R.sub.1A-CO--, where R.sub.1A may be selected, for example, from
the group consisting of a straight or branched alkyl group of 1 to
6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, a
cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl
part thereof and 1 to 3 carbon atoms in the alkyl part thereof, an
alkyloxy group of 1 to 6 carbon atom, --CH.sub.2OH,
CH.sub.3O.sub.2C--, CH.sub.3O.sub.2CCH.sub.2--,
Acetyl-OCH.sub.2CH.sub.2--, HO.sub.2CCH.sub.2--, 2-hydroxyphenyl,
3-hydroxyphenyl, 4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula
##STR29##
[0075] a picolyl group selected from the group consisting of
##STR30##
[0076] a picolyloxy group selected from the group consisting of
##STR31##
[0077] a substituted pyridyl group selected from the group
consisting of ##STR32##
[0078] and a group of formula, ##STR33##
[0079] wherein X', Y', R.sub.4 and R.sub.5 are as defined
herein.
[0080] Exemplary embodiments of Lysine-based compounds encompassed
by the present invention and which may be used in combination with
a CYP450 inhibitor in the methods, pharmaceutical compositions,
kits and uses described herein are also defined by a compound of
formula II, ##STR34##
[0081] pharmaceutically acceptable salts and derivatives thereof
(e.g., for example, when the compound of the present invention
comprises an amino group, the pharmaceutically acceptable salt may
be an ammonium salt),
[0082] wherein n may be 3 or 4,
[0083] wherein X and Y, the same or different, may be selected, for
example, from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5,
--NHCOR.sub.4, --OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4,
and --CH.sub.2OH or X and Y together define an alkylenedioxy group
selected from the group consisting of a methylenedioxy group of
formula --OCH.sub.2O-- and an ethylenedioxy group of formula
--OCH.sub.2CH.sub.2O--,
[0084] wherein R.sub.6 may be selected, for example, from the group
consisting of a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof,
[0085] wherein R.sub.3 may be selected, for example, from the group
consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of
3 to 6 carbon atoms, and a group of formula R.sub.3A-CO--, wherein
R.sub.3A may be selected, for example, from the group consisting of
a straight or branched alkyl group of 1 to 6 carbon atoms (e.g.
methyl, ethyl-, propyl, iso-propyl, butyl, iso-butyl, tert-butyl,
tert-butyl-CH.sub.2--, etc.), a cycloalkyl group having 3 to 6
carbon atoms (e.g. cyclopropyl-, cyclohexyl-etc.), a
cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl
part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
(e.g. cyclopropyl-CH.sub.2--, cyclohexyl-CH.sub.2--, etc.), an
alkyloxy group of 1 to 6 carbon atoms (e.g. CH.sub.3O--,
CH.sub.3CH.sub.2O--, iso-butylO-, tert-butylO-(Boc), etc.),
tetrahydro-3-furanyloxy, --CH.sub.2OH, --CF.sub.3,
--CH.sub.2CF.sub.3, --CH.sub.2CH.sub.2CF.sub.3, pyrrolidinyl,
piperidinyl, 4-morpholinyl, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 3-hydroxyphenyl, 4-hydroxyphenyl,
4-CH.sub.3OC.sub.6H.sub.4CH.sub.2--, CH.sub.3NH--,
(CH.sub.3).sub.2N--, (CH.sub.3CH.sub.2).sub.2N--,
(CH.sub.3CH.sub.2CH.sub.2).sub.2N--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--,
C.sub.6H.sub.5CH.sub.2O--, 2-pyrrolyl, 2-pyridyl, 3-pyridyl,
4-pyridyl, 2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl,
1-isoquinolyl, 3-isoquinolyl, 2-quinoxalinyl, a phenyl group of
formula ##STR35##
[0086] a picolyl group selected from the group consisting of
##STR36##
[0087] a picolyloxy group selected from the group consisting of
##STR37##
[0088] a substituted pyridyl group selected from the group
consisting of ##STR38##
[0089] and a group of formula, ##STR39##
[0090] wherein X' and Y', the same or different, may be selected,
for example, from the group consisting of H, a straight alkyl group
of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon
atoms, a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4 and
--CH.sub.2OH,
[0091] wherein R.sub.4 and R.sub.5, the same or different, may be
selected, for example, from the group consisting of H, a straight
alkyl group of 1 to 6 carbon atoms, a branched alkyl group of 3 to
6 carbon atoms, and a cycloalkyl group of 3 to 6 carbon atoms,
[0092] wherein R.sub.2 may be selected from the group consisting of
a diphenylmethyl group of formula IV ##STR40##
[0093] a naphthyl-1-CH.sub.2-- group of formula V ##STR41##
[0094] a naphthyl-2-CH.sub.2-- group of formula VI ##STR42##
[0095] a biphenylmethyl group of formula VII ##STR43##
[0096] and an anthryl-9-CH.sub.2-- group of formula VIII
##STR44##
[0097] and wherein R.sub.1 may be a physiologically cleavable unit,
whereby upon cleavage of the unit the compound may be able to
release a protease inhibitor, provided that R.sub.1 is not H.
[0098] In accordance with the present invention, R.sub.1 may be
selected, for example, from the group consisting of (HO).sub.2P(O)
and (MO).sub.2P(O), wherein M is an alkali metal (e.g. Na, K, Cs,
etc) or alkaline earth metal (Ca, Mg, etc.).
[0099] Further in accordance with the present invention R.sub.1 may
be a group of formula R.sub.1A--CO--, wherein R.sub.1A may be
selected from the group consisting of a straight or branched alkyl
group of 1 to 6 carbon atoms (e.g. methyl, ethyl, propyl,
iso-propyl, butyl, iso-butyl, tert-butyl, tert-butyl-CH.sub.2--,
etc.), a cycloalkyl group having 3 to 6 carbon atoms (e.g.
cyclopropyl-, cyclohexyl- etc.), a cycloalkylalkyl group having 3
to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon
atoms in the alkyl part thereof, (e.g. cyclopropyl-CH.sub.2--,
cyclohexyl-CH.sub.2--, etc.), an alkyloxy group of 1 to 6 carbon
atoms (e.g. CH.sub.3O--, CH.sub.3CH.sub.2O--, iso-butylO-,
tert-butylO-(Boc), etc.), --CH.sub.2OH, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 2-hydroxyphenyl, 3-hydroxyphenyl,
4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula
##STR45##
[0100] a picolyl group selected from the group consisting of
##STR46##
[0101] a picolyloxy group selected from the group consisting of
##STR47##
[0102] a substituted pyridyl group selected from the group
consisting of ##STR48##
[0103] and a group of formula, ##STR49##
[0104] wherein X', Y', R.sub.4 and R.sub.5 are as defined
herein.
[0105] As mentioned herein, active ingredients disclosed in U.S.
Pat. No. 6,632,816 to Stranix et al., which may be used in
combination with a CYP450 inhibitor in the methods, pharmaceutical
compositions, kits and uses described herein, are defined by a
compound of formula II (or a salt thereof), wherein R.sub.1 is H
and R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, n, X, Y, X', Y'
are as defined herein.
[0106] More particularly, the present invention provides in one
aspect thereof, a pharmaceutical composition which may
comprise;
[0107] a) a compound of formula II and pharmaceutically acceptable
salts thereof,
[0108] b) a cytochrome P450 monooxigenase inhibitor, and;
[0109] c) a pharmaceutically acceptable carrier;
[0110] where the compound of formula II is represented by;
##STR50##
[0111] wherein n may be 3 or 4,
[0112] wherein X and Y, the same or different, may be selected from
the group consisting of H, a straight alkyl group of 1 to 6 carbon
atoms, a branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl
group of 3 to 6 carbon atoms, F, Cl, Br, I, --CF.sub.3,
--OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4, and --CH.sub.2OH
or X and Y together may define, for example, an alkylenedioxy group
which may be selected from the group consisting of a methylenedioxy
group of formula --OCH.sub.2O-- and an ethylenedioxy group of
formula --OCH.sub.2CH.sub.2O--,
[0113] wherein R.sub.6 may be selected, for example, from the group
consisting of a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof,
[0114] wherein R.sub.3 may be selected, for example, from the group
consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a
branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of
3 to 6 carbon atoms, and a group of formula R.sub.3A--CO--, where
R.sub.3A may be selected, for example, from the group consisting of
a straight or branched alkyl group of 1 to 6 carbon atoms, a
cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof, an alkyloxy group of
1 to 6 carbon atoms, tetrahydro-3-furanyloxy, --CH.sub.2OH,
--CF.sub.3, --CH.sub.2CF.sub.3, --CH.sub.2CH.sub.2CF.sub.3,
pyrrolidinyl, piperidinyl, 4-morpholinyl, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 3-hydroxyphenyl, 4-hydroxyphenyl,
4-CH.sub.3OC.sub.6H.sub.4CH.sub.2--, CH.sub.3NH--,
(CH.sub.3).sub.2N--, (CH.sub.3CH.sub.2).sub.2N--,
(CH.sub.3CH.sub.2CH.sub.2).sub.2N--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3CH.sub.2--, CH.sub.3OCH.sub.2CH.sub.2O--,
C.sub.6H.sub.5CH.sub.2O--, 2-pyrrolyl, 2-pyridyl, 3-pyridyl,
4-pyridyl-, 2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl,
1-isoquinolyl, 3-isoquinolyl, 2-quinoxalinyl, a phenyl group of
formula ##STR51##
[0115] a picolyl group which may be selected from the group
consisting of ##STR52##
[0116] a picolyloxy group which may be selected from the group
consisting of ##STR53##
[0117] a substituted pyridyl group which may be selected from the
group consisting of ##STR54##
[0118] a group of formula ##STR55##
[0119] wherein X' and Y', the same or different, may be selected
from the group consisting of H, a straight alkyl group of 1 to 6
carbon atoms, a branched alkyl group of 3 to 6 carbon atoms, a
cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I, --CF.sub.3,
--NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4, --OR.sub.4,
--SR.sub.4, --COOR.sub.4, --COR.sub.4 and --CH.sub.2OH,
[0120] wherein R.sub.4 and R.sub.5, the same or different, may be
selected from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
and a cycloalkyl group of 3 to 6 carbon atoms,
[0121] wherein R.sub.2 may be selected from the group consisting of
a diphenylmethyl group of formula IV ##STR56##
[0122] a naphthyl-1-CH.sub.2-- group of formula V ##STR57##
[0123] a naphthyl-2-CH.sub.2-- group of formula VI ##STR58##
[0124] a biphenylmethyl group of formula VII ##STR59##
[0125] and an anthryl-9-CH.sub.2-- group of formula VIII
##STR60##
[0126] wherein R.sub.1 may be H or a physiologically cleavable
unit, whereby upon physiological conditions (in vivo) the compound
may be converted into an active protease inhibitor.
[0127] In accordance with the present invention, R.sub.1 may be
selected, for example, from the group consisting of H,
(HO).sub.2P(O) and (MO).sub.2P(O) (wherein M may be an alkali metal
or alkaline earth metal), and a group of formula R.sub.1A--CO--,
where R.sub.1A may be selected, for example, from the group
consisting of a straight or branched alkyl group of 1 to 6 carbon
atoms, a cycloalkyl group having 3 to 6 carbon atoms, a
cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl
part thereof and 1 to 3 carbon atoms in the alkyl part thereof, an
alkyloxy group of 1 to 6 carbon atoms, --CH.sub.2OH,
CH.sub.3O.sub.2C--, CH.sub.3O.sub.2CCH.sub.2--,
Acetyl-OCH.sub.2CH.sub.2--, HO.sub.2CCH.sub.2--, 2-hydroxyphenyl,
3-hydroxyphenyl, 4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula
##STR61##
[0128] a picolyl group which may be selected from the group
consisting of ##STR62##
[0129] a picolyloxy group which may be selected from the group
consisting of ##STR63##
[0130] a substituted pyridyl group which may be selected from the
group consisting of ##STR64##
[0131] and a group of formula, ##STR65##
[0132] wherein X', Y', R.sub.4 and R.sub.5 are as defined
herein.
[0133] In accordance with an embodiment of the present invention,
pharmaceutical compositions which comprise compounds of formula II
wherein R.sub.6 may be, for example, iso-butyl and n may be 3 are
encompassed herewith.
[0134] In accordance with an additional embodiment of the present
invention, pharmaceutical compositions which comprise compounds of
formula II wherein R.sub.6 may be, for example, iso-butyl and n may
be 4 are also encompassed herewith.
[0135] In a particular embodiment of the present invention, R.sub.1
may be selected, for example, from the group consisting of H,
(HO).sub.2P(O) and (NaO).sub.2P(O).
[0136] In another particular embodiment of the present invention,
R.sub.1 may be selected, for example, from the group consisting of
CH.sub.3CO, 3-pyridyl-CO, (CH.sub.3).sub.2NCH.sub.2CO and
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
[0137] In accordance with another particular embodiment of the
present invention, pharmaceutical compositions which comprise
compounds of formula II wherein R.sub.3 may be selected, for
example, from the group consisting of CH.sub.3CO, CH.sub.3O--CO,
(CH.sub.3).sub.2N--CO, 3-pyridyl-CO, 4-pyridyl-CO and
4-morpholine-CO are encompassed by the present invention.
[0138] In accordance with an embodiment of the present invention, X
may be 4-NH.sub.2 and Y may be H or F.
[0139] In accordance with an embodiment of the present invention,
X' and Y' may both be H.
[0140] In accordance with a particular embodiment of the present
invention, pharmaceutical compositions which comprise compounds of
formula II wherein R.sub.2 may be selected, for example, from the
group consisting of a diphenylmethyl group of formula IV, a
naphthyl-1-CH.sub.2-- group of formula V, a naphthyl-2-CH.sub.2--
group of formula VI, a biphenylmethyl group of formula VII and an
anthryl-9-CH.sub.2-- group of formula VIII are encompassed
herewith.
[0141] For example, R.sub.2 may, more particularly, be selected
from the group consisting of a diphenylmethyl group of formula IV,
a naphthyl-1-CH.sub.2-- group of formula V, and a
naphthyl-2-CH.sub.2-- group of formula VI.
[0142] In a further aspect, the present invention provides
pharmaceutical compositions comprising a compound of formula II,
wherein R.sub.6 is isobutyl, n is 4 and R.sub.2 is a diphenylmethyl
group of formula IV.
[0143] In accordance with an embodiment of the present invention,
R.sub.1 may be selected from the group consisting of H,
(HO).sub.2P(O) and (NaO).sub.2P(O).
[0144] In accordance with a further embodiment of the present
invention, R.sub.1 may be selected from the group consisting of
CH.sub.3CO, 3-pyridyl-CO, (CH.sub.3).sub.2NCH.sub.2CO and
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
[0145] In accordance with an additional embodiment of the present
invention, R.sub.3 may be selected, for example, from the group
consisting of CH.sub.3CO, CH.sub.3O--CO, (CH.sub.3).sub.2N--CO,
3-pyridyl-CO, 4-pyridyl-CO and 4-morpholine-CO.
[0146] In accordance with an embodiment of the present invention, X
may be 4-NH.sub.2 and Y may be H or F.
[0147] More particularly, in accordance with an embodiment thereof,
the present invention provides a pharmaceutical composition which
may comprise a compound of formula II, wherein X may be, for
example, 4-NH.sub.2, Y may be H, X' may be H, Y' may be H and
R.sub.3 may be CH.sub.3O--CO.
[0148] In accordance with a particular embodiment of the present
invention, R.sub.1 may be (HO).sub.2P(O).
[0149] In accordance with another particular embodiment of the
present invention, R.sub.1 may be (NaO).sub.2P(O).
[0150] In accordance with a further particular embodiment of the
present invention, R.sub.1 may be H.
[0151] More particularly, in accordance with a further embodiment
thereof, the present invention provides a pharmaceutical
composition which may comprise a compound of formula II, wherein X
may be 4-NH.sub.2, Y may be 3-F, X' may be H, Y' may be H and
R.sub.3 may be CH.sub.3O--CO.
[0152] In accordance with a particular embodiment of the present
invention, R.sub.1 may be (HO).sub.2P(O).
[0153] In accordance with another particular embodiment of the
present invention, R.sub.1 may be (NaO).sub.2P(O).
[0154] In accordance with a further particular embodiment of the
present invention, R.sub.1 may be H.
[0155] More particularly, in accordance with an additional
embodiment thereof, the present invention provides a pharmaceutical
composition which may comprise a compound of formula II, wherein X
is 4-NH.sub.2, Y is H or 3-F, X' is H, Y' is H and R.sub.3 is
CH.sub.3CO.
[0156] In accordance with a particular embodiment of the present
invention, R.sub.1 may be (HO).sub.2P(O).
[0157] In accordance with another particular embodiment of the
present invention, R.sub.1 may be (NaO).sub.2P(O).
[0158] In accordance with a further particular embodiment of the
present invention, R.sub.1 may be H.
[0159] More particularly, in accordance with an embodiment thereof,
the present invention further provides a pharmaceutical composition
which may comprise a compound of formula II, X is 4-NH.sub.2, Y is
H or 3-F, X' is H, Y' is H and R.sub.3 is 4-morpholine-CO.
[0160] In accordance with an embodiment of the present invention X
may be 4-NH.sub.2, Y may be H, X' may be H, Y' may be H and R.sub.3
may be CH.sub.3O--CO.
[0161] In accordance with a particular embodiment of the present
invention R.sub.1 may be 3-pyridyl-CO.
[0162] In accordance with another particular embodiment of the
present invention R.sub.1 may be (CH.sub.3).sub.2NCH.sub.2CO.
[0163] In accordance with yet another particular embodiment of the
present invention R.sub.1 may be
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
[0164] In accordance with an additional particular embodiment of
the present invention R.sub.1 may be CH.sub.3CO.
[0165] In an additional embodiment the present invention provides
pharmaceutical compositions comprising a compound of formula II,
wherein R.sub.6 is iso-butyl, n is 4, X' and Y' are both H, R.sub.2
is Naphtyl-1-CH.sub.2--, X is 4-NH.sub.2, Y is H, R.sub.3 is
4-morpholine-CO and R.sub.1 may be selected, for example, from the
group consisting of H, (HO).sub.2P(O) and (NaO).sub.2P(O).
[0166] In a further embodiment, the present invention provides
pharmaceutical compositions comprising a compound of formula II,
wherein R.sub.6 is iso-butyl, n is 4, X' and Y' are both H, R.sub.2
is Naphtyl-2-CH.sub.2--, X is 4-NH.sub.2, Y is H, R.sub.3 is
CH.sub.3O--CO and R.sub.1 may be selected, for example, from the
group consisting of H, (HO).sub.2P(O) and (NaO).sub.2P(O).
[0167] In accordance with the present invention, the cytochrome
P450 monooxigenase inhibitor may be a CYP450-3A inhibitor.
[0168] In accordance with the present invention, the cytochrome
P450 monooxigenase inhibitor may be selected, for example, from the
group consisting of ritonavir (RTV), ketoconazole, fluconazole,
nefazodone, fluvoxamine, fluoxetine, macrolide antibiotics,
sertraline sulfaphenazole and erythromycin.
[0169] Also in accordance with the present invention, the
pharmaceutical composition may be administered, for example,
orally.
[0170] Further in accordance with the present invention, the
pharmaceutical composition may be administered, for example,
twice-daily.
[0171] In accordance with an embodiment of the present invention,
the ratio of (the concentration of the) compound of formula I over
the (concentration of the) cytochrome P450 monooxygenase inhibitor
may be between about 1 (e.g., 1:1) to about 10 (e.g., 10:1).
[0172] Also in accordance with an embodiment of the present
invention, the ratio of compound of formula I over the cytochrome
P450 monooxygenase inhibitor may be, for example, between about 3:1
and about 6:1.
[0173] Further exemplary embodiments of Lysine-based compounds
which may be used to carry out the present invention may include,
for example, a compound of formula IIa; ##STR66##
[0174] pharmaceutically acceptable salts and derivatives thereof
(e.g., for example, when the compound of the present invention
comprises an amino group, the pharmaceutically acceptable salt may
be an ammonium salt),
[0175] wherein X and Y, the same or different, may be selected, for
example, from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5,
--NHCOR.sub.4, --OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4,
and --CH.sub.2OH or X and Y together define an alkylenedioxy group
selected from the group consisting of a methylenedioxy group of
formula --OCH.sub.2O-- and an ethylenedioxy group of formula
--OCH.sub.2CH.sub.2O--,
[0176] wherein X' and Y', the same or different, may be selected,
for example, from the group consisting of H, a straight alkyl group
of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon
atoms, a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4 and
--CH.sub.2OH,
[0177] and wherein n, R.sub.1, R.sub.3, R.sub.4, R.sub.5 and
R.sub.6 are as defined herein.
[0178] Active ingredients disclosed in U.S. Pat. No. 6,632,816 to
Stranix et al, which may be used in combination with a CYP450
inhibitor in the methods, pharmaceutical compositions, kits and
uses described herein, are defined by a compound of formula IIa (or
a salt thereof), wherein R.sub.1 is H and R.sub.3, R.sub.4,
R.sub.5, R.sub.6, n, X, Y, X', Y' are as defined herein.
[0179] More particularly, the present invention provides a
pharmaceutical composition which may comprise;
[0180] a) a compound of formula IIa ##STR67##
[0181] and pharmaceutically acceptable salts thereof,
[0182] wherein X and Y, the same or different, may be selected, for
example, from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5,
--NHCOR.sub.4, --OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4,
and --CH.sub.2OH or X and Y together define an alkylenedioxy group
selected from the group consisting of a methylenedioxy group of
formula --OCH.sub.2O-- and an ethylenedioxy group of formula
--OCH.sub.2CH.sub.2O--,
[0183] wherein X' and Y', the same or different, may be selected,
for example, from the group consisting of H, a straight alkyl group
of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon
atoms, a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4 and
--CH.sub.2OH,
[0184] wherein n, R.sub.1, R.sub.3, R.sub.4, R.sub.5 and R.sub.6
are as defined herein;
[0185] b) a cytochrome P450 monooxigenase inhibitor, and;
[0186] c) a pharmaceutically acceptable carrier.
[0187] In accordance with an embodiment of the present invention,
R.sub.1 may be selected from the group consisting of H,
(HO).sub.2P(O) and (MO).sub.2P(O), wherein M may be an alkali metal
or alkaline earth metal and a group of formula R.sub.1A--CO--,
R.sub.1A which may be selected from the group consisting of a
straight or branched alkyl group of 1 to 6 carbon atoms, a
cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof, an alkyloxy group of
1 to 6 carbon atom, --CH.sub.2OH, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 2-hydroxyphenyl, 3-hydroxyphenyl,
4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula
##STR68##
[0188] a picolyl group selected from the group consisting of
##STR69##
[0189] a picolyloxy group selected from the group consisting of
##STR70##
[0190] a substituted pyridyl group selected from the group
consisting of ##STR71##
[0191] and a group of formula, ##STR72##
[0192] wherein X', Y', R.sub.4 and R.sub.5 are as defined
herein.
[0193] In accordance with another embodiment of the present
invention, R.sub.6 may be iso-butyl.
[0194] In accordance with yet another embodiment of the present
invention, n may be 4.
[0195] In accordance with a further embodiment of the present
invention, R.sub.1 may be selected from the group consisting of H,
(HO).sub.2P(O) and (NaO).sub.2P(O).
[0196] Further in accordance with an embodiment of the present
invention, R.sub.1 may be selected, for example, from the group
consisting of CH.sub.3CO, 3-pyridyl-CO, (CH.sub.3).sub.2NCH.sub.2CO
and (CH.sub.3).sub.2CHCH(NH.sub.2)CO.
[0197] Also in accordance with an embodiment of the present
invention, R.sub.3 may be selected from the group consisting of
CH.sub.3CO, CH.sub.3O--CO, (CH.sub.3).sub.2N--CO, 3-pyridyl-CO,
4-pyridyl-CO and 4-morpholine-CO.
[0198] In accordance with another embodiment of the present
invention, R.sub.3 may be selected from the group consisting of
CH.sub.3CO, CH.sub.3O--CO, (CH.sub.3).sub.2N--CO, 3-pyridyl-CO,
4-pyridyl-CO and 4-morpholine-CO.
[0199] In accordance with an embodiment of the present invention, X
may be 4-NH.sub.2 and Y may be H or F.
[0200] In accordance with another embodiment of the present
invention, X may be 4-NH.sub.2 and Y may be H or F.
[0201] Also in accordance with an embodiment of the present
invention, X may be 4-NH.sub.2, Y may be H or 3-F, X' may be H, Y'
may be H and R.sub.3 may be CH.sub.3CO.
[0202] In accordance with another embodiment of the present
invention, X may be 4-NH.sub.2, Y may be H or 3-F, X' may be H, Y'
may be H and R.sub.3 may be 4-morpholine-CO.
[0203] In accordance with a particular embodiment of the present
invention, X may be 4-NH.sub.2, Y may be H, X' may be H, Y' may be
H, R.sub.3 may be CH.sub.3O--CO and R.sub.1 may be (HO).sub.2P(O),
(NaO).sub.2P(O) or H.
[0204] In accordance with another particular embodiment of the
present invention, X may be 4-NH.sub.2, Y may be 3-F, X' may be H,
Y' may be H, R.sub.3 may be CH.sub.3O--CO and R.sub.1 may be
(HO).sub.2P(O), (NaO).sub.2P(O) or H.
[0205] In accordance with a further particular embodiment of the
present invention, X may be 4-NH.sub.2, Y may be H or 3-F, X' may
be H, Y' may be H, R.sub.3 may be CH.sub.3CO and R.sub.1 may be
(HO).sub.2P(O), (NaO).sub.2P(O) or H.
[0206] In accordance with another particular embodiment of the
present invention, X may be 4-NH.sub.2, Y may be H or 3-F, X' may
be H, Y' may be H and R.sub.3 may be 4-morpholine-CO.
[0207] In accordance with an additional embodiment of the present
invention, X may be 4-NH.sub.2, Y may be H, X' may be H, Y' may be
H, R.sub.3 may be CH.sub.3O--CO and R.sub.1 may be 3-pyridyl-CO,
(CH.sub.3).sub.2NCH.sub.2CO, (CH.sub.3).sub.2CHCH(NH.sub.2)CO or
CH.sub.3CO.
[0208] In accordance with another embodiment of the present
invention, X may be 4-NH.sub.2, Y may be 3-F, X' may be H, Y' may
be H, R.sub.3 may be CH.sub.3O--CO and R.sub.1 may be 3-pyridyl-CO,
(CH.sub.3).sub.2NCH.sub.2CO or
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
[0209] Additional embodiments of Lysine-based compounds which may
be used to carry out the present invention include, a compound of
formula IIb ##STR73##
[0210] pharmaceutically acceptable salts and derivatives thereof
(e.g., for example, when the compound of the present invention
comprises an amino group, the pharmaceutically acceptable salt may
be an ammonium salt),
[0211] wherein X and Y, the same or different, may be selected, for
example, from the group consisting of H, a straight alkyl group of
1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms,
a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5,
--NHCOR.sub.4, --OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4,
and --CH.sub.2OH or X and Y together define an alkylenedioxy group
selected from the group consisting of a methylenedioxy group of
formula --OCH.sub.2O-- and an ethylenedioxy group of formula
--OCH.sub.2CH.sub.2O--,
[0212] wherein X' and Y', the same or different, may be selected,
for example, from the group consisting of H, a straight alkyl group
of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon
atoms, a cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I,
--CF.sub.3, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4 and --CH.sub.2OH,
and wherein n, R.sub.1, R.sub.3, R.sub.4, R.sub.5 and R.sub.6 are
as defined herein.
[0213] Active ingredients disclosed in U.S. Pat. No. 6,632,816 to
Stranix et al, which may be used in combination with a CYP450
inhibitor in the methods, pharmaceutical compositions, kits and
uses described herein, are defined by a compound of formula IIb (or
a salt thereof), wherein R.sub.1 is H and R.sub.3, R.sub.4,
R.sub.5, R.sub.6, n, X, Y, X', Y' are as defined herein.
[0214] The present invention therefore, more particularly provides
a pharmaceutical composition which may comprise,
[0215] a) a compound of formula IIb ##STR74##
[0216] and pharmaceutically acceptable salts thereof,
[0217] wherein X and Y, the same or different, may be selected from
the group consisting of H, a straight alkyl group of 1 to 6 carbon
atoms, a branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl
group of 3 to 6 carbon atoms, F, Cl, Br, I, --CF.sub.3,
--OCF.sub.3, --CN, --NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4,
--OR.sub.4, --SR.sub.4, --COOR.sub.4, --COR.sub.4, and --CH.sub.2OH
or X and Y together define an alkylenedioxy group which may be
selected from the group consisting of a methylenedioxy group of
formula --OCH.sub.2O-- and an ethylenedioxy group of formula
--OCH.sub.2CH.sub.2O--,
[0218] wherein X' and Y', the same or different, may be selected
from the group consisting of H, a straight alkyl group of 1 to 6
carbon atoms, a branched alkyl group of 3 to 6 carbon atoms, a
cycloalkyl group of 3 to 6 carbon atoms, F, Cl, Br, I, --CF.sub.3,
--NO.sub.2, --NR.sub.4R.sub.5, --NHCOR.sub.4, --OR.sub.4,
--SR.sub.4, --COOR.sub.4, --COR.sub.4 and --CH.sub.2OH,
[0219] wherein n, R.sub.1, R.sub.3, R.sub.4, R.sub.5 and R.sub.6
are as defined herein;
[0220] b) a cytochrome P450 monooxigenase inhibitor, and;
[0221] c) a pharmaceutically acceptable carrier.
[0222] In accordance with the present invention, R.sub.1 may be
selected from the group consisting of H, (HO).sub.2P(O) and
(MO).sub.2P(O), wherein M may be an alkali metal or alkaline earth
metal and a group of formula R.sub.1A--CO--, R.sub.1A which may be
selected from the group consisting of a straight or branched alkyl
group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6
carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in
the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl
part thereof, an alkyloxy group of 1 to 6 carbon atom,
--CH.sub.2OH, CH.sub.3O.sub.2C--, CH.sub.3O.sub.2CCH.sub.2--,
Acetyl-OCH.sub.2CH.sub.2--, HO.sub.2CCH.sub.2--, 2-hydroxyphenyl,
3-hydroxyphenyl, 4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula
##STR75##
[0223] a picolyl group selected from the group consisting of
##STR76##
[0224] a picolyloxy group selected from the group consisting of
##STR77##
[0225] a substituted pyridyl group selected from the group
consisting of ##STR78##
[0226] and a group of formula; ##STR79##
[0227] wherein X', Y', R.sub.4 and R.sub.5 are as defined
herein.
[0228] In accordance with an embodiment of the present invention,
R.sub.6 may be iso-butyl.
[0229] In accordance with another embodiment of the present
invention, n may be 4.
[0230] In accordance with a further embodiment of the present
invention, R.sub.1 may be selected, for example, from the group
consisting of H, (HO).sub.2P(O) and (NaO).sub.2P(O).
[0231] In accordance with an additional embodiment of the present
invention, R.sub.1 may selected, for example, from the group of
CH.sub.3CO, 3-pyridyl-CO, (CH.sub.3).sub.2NCH.sub.2CO and
(CH.sub.3).sub.2CHCH(NH.sub.2)CO.
[0232] In accordance with yet a further embodiment of the present
invention, R.sub.3 may be selected, for example, from the group
consisting of CH.sub.3O--CO, (CH.sub.3).sub.2N--CO, 3-pyridyl-CO,
4-pyridyl-CO and 4-morpholine-CO.
[0233] In accordance with an embodiment of the present invention,
R.sub.3 may be selected, for example, from the group consisting of
CH.sub.3O--CO, (CH.sub.3).sub.2N--CO, 3-pyridyl-CO, 4-pyridyl-CO
and 4-morpholine-CO.
[0234] Also in accordance with an embodiment of the present
invention, X may be, for example, 4-NH.sub.2 and Y may be H or
F.
[0235] Further in accordance with an embodiment of the present
invention, X may be, for example, 4-NH.sub.2 and Y may be H or
F.
[0236] In accordance with a particular embodiment of the present
invention, X may be 4-NH.sub.2, Y may be H or 3-F, X' may be H, Y'
may be H and R.sub.3 may be CH.sub.3O--CO.
[0237] In accordance with a further particular embodiment of the
present invention, X may be 4-NH.sub.2, Y may be H or 3-F, X' may
be H, Y' may be H and R.sub.3 may be CH.sub.3CO.
[0238] In accordance with another particular embodiment of the
present invention, X may be 4-NH.sub.2, Y may be H or 3-F, X' may
be H, Y' may be H and R.sub.3 may be 4-morpholine-CO.
[0239] In accordance with an embodiment of the present invention
the naphthyl group may be, for example, a naphthyl-2-CH.sub.2
group, Y may be H and R.sub.1 may be (HO).sub.2P(O).
[0240] In accordance with another embodiment of the present
invention the naphthyl group may be, for example, a
naphthyl-1-CH.sub.2 group. Y may be H and R.sub.1 may be
(HO).sub.2P(O).
[0241] Further exemplary embodiments of compounds which may be used
in the present invention, include, for example, a compound of
formula llc ##STR80##
[0242] pharmaceutically acceptable salts and derivatives thereof
(e.g., for example, when the compound of the present invention
comprises an amino group, the pharmaceutically acceptable salt may
be an ammonium salt),
[0243] and wherein n, X, Y, X', Y', R.sub.1, R.sub.3, R.sub.4,
R.sub.5 and R.sub.6 are as defined herein.
[0244] Active ingredients disclosed in U.S. Pat. No. 6,632,816 to
Stranix et al., which may be used in combination with a CYP450
inhibitor in the methods, pharmaceutical compositions, kits and
uses described herein, are defined by a compound of formula IIc (or
a salt thereof), wherein R.sub.1 is H and R.sub.3, R.sub.4,
R.sub.5, R.sub.6, n, X, Y, X', Y' are as defined herein.
[0245] The present invention more particularly provides a
pharmaceutical composition which may comprise
[0246] a) a compound of formula IIc ##STR81##
[0247] and pharmaceutically acceptable salts thereof,
[0248] wherein n, X, Y, X', Y', R.sub.1, R.sub.3, R.sub.4, R.sub.5
and R.sub.6 are as defined herein;
[0249] b) a cytochrome P450 monooxigenase inhibitor, and;
[0250] c) a pharmaceutically acceptable carrier.
[0251] In accordance with an embodiment of the present invention,
R.sub.1 may be selected, for example, from the group consisting of
H, (HO).sub.2P(O) and (MO).sub.2P(O), wherein M may be an alkali
metal or alkaline earth metal and a group of formula R.sub.1A-CO--,
R.sub.1A which may be selected from the group consisting of a
straight or branched alkyl group of 1 to 6 carbon atoms, a
cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl
group having 3 to 6 carbon atoms in the cycloalkyl part thereof and
1 to 3 carbon atoms in the alkyl part thereof, an alkyloxy group of
1 to 6 carbon atom, --CH.sub.2OH, CH.sub.3O.sub.2C--,
CH.sub.3O.sub.2CCH.sub.2--, Acetyl-OCH.sub.2CH.sub.2--,
HO.sub.2CCH.sub.2--, 2-hydroxyphenyl, 3-hydroxyphenyl,
4-hydroxyphenyl, (CH.sub.3).sub.2NCH.sub.2--,
(CH.sub.3).sub.2CHCH(NH.sub.2)--, HOCH.sub.2CH.sub.2NH--,
CH.sub.3OCH.sub.2O--, CH.sub.3OCH.sub.2CH.sub.2O--, 2-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 1-methyl-1,4-dihydro-3-pyridyl,
2-pyrazinyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 1-isoquinolyl,
3-isoquinolyl, 2-quinoxalinyl, a phenyl group of formula
##STR82##
[0252] a picolyl group selected from the group consisting of
##STR83##
[0253] a picolyloxy group selected from the group consisting of
##STR84##
[0254] a substituted pyridyl group selected from the group
consisting of ##STR85##
[0255] and a group of formula, ##STR86##
[0256] wherein X', Y', R.sub.4 and R.sub.5 are as defined
herein.
[0257] In accordance with another embodiment of the present
invention, R.sub.6 may be iso-butyl.
[0258] In accordance with yet another embodiment of the present
invention, n may be 4.
[0259] In accordance with a further embodiment of the present
invention, R.sub.1 may be, for example, H(HO).sub.2P(O) or
(NaO).sub.2P(O).
[0260] In accordance with an additional embodiment of the present
invention, R.sub.1 may be selected, for example, from the group
consisting of CH.sub.3CO, 3-pyridyl-CO, (CH.sub.3).sub.2NCH.sub.2CO
and (CH.sub.3).sub.2CHCH(NH.sub.2)CO.
[0261] Other compounds which may be used to carry out the present
invention may include, for example, a compound of formula IIA;
##STR87##
[0262] wherein Y, n, R.sub.1, R.sub.2, R.sub.3, X' and Y' are as
defined herein.
[0263] In accordance with the present invention, R.sub.1 may be,
for example, (HO).sub.2P(O) or (NaO).sub.2P(O). Further in
accordance with the present invention, n may be 4. Y may be, for
example, H. R.sub.3 may be, for example, CH.sub.3O--CO. R.sub.2 may
be, for example, a diphenylmethyl group of formula IV, where X' and
Y' may be, for example H, ##STR88##
[0264] Active ingredients disclosed in U.S. Pat. No. 6,632,816 to
Stranix et al, which may be used in combination with a CYP450
inhibitor in the methods, pharmaceutical compositions, kits and
uses described herein, are defined by a compound of formula IIA (or
a salt thereof), wherein R.sub.1 is H and R.sub.2, R.sub.3,
R.sub.4, R.sub.5, n, X, Y, X', Y' are as defined herein.
[0265] Compounds of formula IIA' as well as pharmaceutically
acceptable salts and derivatives thereof may be used to carry out
the present invention, ##STR89##
[0266] such as, for example, compound of formula IIA' wherein
R.sub.1 is (HO).sub.2P(O) or, compound of formula IIA' wherein
R.sub.1 is (NaO).sub.2P(O).
[0267] Active ingredients disclosed in U.S. Pat. No. 6,632,816 to
Stranix et al, which may be used in combination with a CYP450
inhibitor in the methods, pharmaceutical compositions, kits and
uses described herein, are defined by a compound of formula IIA'
(or a salt thereof), wherein R.sub.1 is H are as defined
herein.
[0268] For example, pharmaceutical compositions, methods, uses and
kits encompassed by the present invention may comprise one or more
of the following compounds and combination thereof;
[0269] a compound of formula IIa wherein n is 4, R.sub.1 is
(HO).sub.2P(O), X is 4-NH.sub.2, Y is H, X'is H, Y' is H, R.sub.6
is iso-butyl and R.sub.3 is CH.sub.3O--CO,
[0270] a compound of formula IIa wherein n is 4, R.sub.1 is
(NaO).sub.2P(O), X is 4-NH.sub.2, Y is H, X' is H, Y' is H, R.sub.6
is iso-butyl and R.sub.3 is CH.sub.3O--CO,
[0271] a compound of formula IIa wherein n is 4, R.sub.1 is
(HO).sub.2P(O), X is 4-NH.sub.2, Y is H, X' is H, Y' is H, R.sub.6
is iso-butyl and R.sub.3 is CH.sub.3CO,
[0272] a compound of formula IIa wherein n is 4, R.sub.1 is
(HO).sub.2P(O), X is 4-NH.sub.2, Y is 3-F, X' is H, Y' is H,
R.sub.6 is iso-butyl and R.sub.3 is CH.sub.3O--CO,
[0273] a compound of formula IIa wherein n is 4, R.sub.1 is
CH.sub.3CO, X is 4-NH.sub.2, Y is H, X' is H, Y' is H, R.sub.6 is
iso-butyl and R.sub.3 is CH.sub.3O--CO,
[0274] a compound of formula IIa wherein n is 4, R.sub.1 is
3-pyridyl-CO, X is 4-NH.sub.2, Y is H, X' is H, Y' is H, R.sub.6 is
iso-butyl and R.sub.3 is CH.sub.3O--CO,
[0275] a compound of formula IIa wherein n is 4, R.sub.1 is
(CH.sub.3).sub.2NCH.sub.2CO, X is 4-NH.sub.2, Y is H, X' is H, Y'
is H, R.sub.6 is iso-butyl and R.sub.3 is CH.sub.3O--CO,
[0276] a compound of formula IIa wherein n is 4, R.sub.1 is
(CH.sub.3).sub.2CHCH(NH.sub.2)CO, X is 4-NH.sub.2, Y is H, X' is H,
Y' is H, R.sub.6 is iso-butyl and R.sub.3 is CH.sub.3O--CO,
[0277] a compound of formula IIb wherein n is 4, R.sub.1 is
(HO).sub.2P(O), X is 4-NH.sub.2, Y is H, X' is H, Y' is H, R.sub.6
is iso-butyl and R.sub.3 is CH.sub.3O--CO and wherein the naphthyl
group is a naphthyl-2-CH.sub.2 group,
[0278] a compound of formula IIb wherein n is 4, R.sub.1 is
(HO).sub.2P(O), X is 4-NH.sub.2, Y is H, X' is H, Y' is H, R.sub.6
is iso-butyl and R.sub.3 is 4-morpholine-CO and wherein the
naphthyl group is a naphthyl-1-CH.sub.2 group, or
[0279] a combination of any of the above mentioned compounds.
[0280] Any compound which is a precursor of an active ingredient
described herein may be used to carry out the present invention and
is also encompassed by the present invention. For example,
compounds being able to release or generate (either in vivo or in
vitro) an active ingredient of the following formula; ##STR90##
[0281] are encompassed by the present invention.
[0282] The above identified active ingredient is identified herein
as PL-100 and it is to be understood herein that any precursor able
to release or generate the above mentioned exemplary compound
either in vivo or in vitro is encompassed by the present invention
and may be used to carry out methods, pharmaceutical compositions,
kits and uses described herein.
[0283] In addition, compounds which able to release (either in vivo
or in vitro) an active ingredient of the following formula;
##STR91##
[0284] may be used to carry out the present invention and are
therefore encompassed by the present invention.
[0285] The above identified active ingredient is identified herein
as PL-337 and it is to be understood herein that any precursor able
to release or generate the above mentioned exemplary compound
either in vivo or in vitro may be used to carry out the present
invention and is encompassed by the present invention.
[0286] The present invention also relates in an additional aspect
thereof, to the use of at least one compound of formula I
##STR92##
[0287] or a pharmaceutically acceptable salts thereof and a
cytochrome P450 monooxigenase inhibitor (e.g., a compound of
formula I in combination with a cytochrome P450 monooxigenase
inhibitor) in the manufacture of a drug for the treatment or
prevention of an HIV infection or for treatment or prevention of
AIDS, wherein n, X, Y, X', Y', R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5 and R.sub.6 are as defined herein.
[0288] In accordance with an embodiment of the present invention,
the cytochrome P450 monooxigenase inhibitor may be selected from
the group consisting of ritonavir (RTV), ketoconazole, fluconazole,
nefazodone, fluvoxamine, fluoxetine, macrolide antibiotics,
sertraline sulfaphenazole and erythromycin.
[0289] The present invention also provides in an additional aspect
thereof to a kit for treating or preventing an HIV infection or for
treating or preventing AIDS, the kit may comprise, for example, a)
a first container which may contain a compound of formula I (e.g.,
in the form of a pharmaceutical composition comprising a compound
of formula I or a pharmaceutically acceptable salts thereof and a
pharmaceutically acceptable carrier); ##STR93##
[0290] wherein n, X, Y, X', Y', R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5 and R.sub.6 are as defined in claim 1, and a second
container containing a cytochrome P450 monooxigenase inhibitor
(e.g., in the form of a pharmaceutical composition comprising a
CYP450 inhibitor and a pharmaceutically acceptable carrier) or;
[0291] b) a container comprising both the compound of formula I and
the CYP450 inhibitor (e.g., in the form of a pharmaceutical
composition comprising a compound of formula I, a CYP450 inhibitor
and a pharmaceutically acceptable carrier).
[0292] In yet an additional aspect, the present invention provides
a method of treating or preventing an HIV infection or of treating
or preventing AIDS, the method may comprise administering a
pharmaceutical composition as defined in claim 1 to a mammal in
need thereof.
[0293] In accordance with an embodiment of the present invention,
administration of the pharmaceutical composition may be performed,
for example, orally.
[0294] In accordance with an embodiment of the present invention
the pharmaceutical composition may be administered, for example,
twice-daily.
[0295] In another aspect, the present invention provides a method
of treating or preventing an HIV infection or of treating or
preventing AIDS, the method may comprise administering (e.g.,
co-administering)
[0296] a) a compound of formula I ##STR94##
[0297] or a pharmaceutically acceptable salts thereof, wherein n,
X, Y, X', Y', R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5 and
R.sub.6 are as defined herein, and;
[0298] b) one or more CYP450 inhibitor in an amount which is
sufficient to reduce the metabolism of the compound of formula
I.
[0299] Further in accordance with the present invention,
administration of the compound of formula I and the CYP450
inhibitor may be performed, for example, separately, simultaneously
or sequentially.
[0300] In accordance with a particular embodiment of the present
invention, administration of the compound of formula I and the
CYP450 inhibitor may be performed, for example, by administering a)
a first pharmaceutical composition which may comprise a compound of
formula I and a pharmaceutically acceptable carrier and b) a second
pharmaceutical composition which may comprise a CYP450 inhibitor
and a pharmaceutically acceptable carrier.
[0301] Further in accordance with the present invention,
administration of the compound of formula I and the CYP450
inhibitor may be performed, for example, by administering a single
pharmaceutical composition which may comprise a compound of formula
I, a CYP450 inhibitor and a pharmaceutically acceptable
carrier.
[0302] The present invention also relates in a further aspect
thereof to a method for improving the pharmacokinetics of a
compound of formula I as defined herein, the method may comprise
administering to a human in need thereof, the compound of formula I
and (e.g., in combination with) an amount of a CYP450 inhibitor
effective to inhibit cytochrome P450 monooxygenase.
[0303] In yet a further aspect, the present invention relates to a
method of inhibiting (e.g., reducing replication of) an HIV (e.g.,
one or more isolate, one or more strain) having a reduced
susceptibility to a protease inhibitor other than the protease
inhibitor defined by formula I, the method may comprise
administering a compound of formula I alone or in combination with
a CYP450 inhibitor to an individual in need thereof.
[0304] In accordance with an embodiment of the present invention,
the HIV may be an HIV-1.
[0305] Also in accordance with an embodiment of the present
invention, the HIV may be one that has a reduced susceptibility to
one or more of a protease inhibitor which may be selected, for
example, from the group consisting of Atazanavir, Amprenavir,
Indinavir, Lopinavir, Nelfinavir, Ritonavir and Saquinavir.
[0306] Further in accordance with an embodiment of the present
invention, the HIV-1 may be one that possesses an aspartyl protease
having one or more mutations (a mutation conferring a resistance to
one or more of protease inhibitor mentioned herein.
[0307] The compounds listed herein are exemplary embodiments of
compounds which may be used to carry out the present invention and
it is to be understood that the present invention is not restricted
to these compounds only.
[0308] The term "pharmaceutically effective amount" refers to an
amount effective in treating, preventing or reducing the risk or
probability of HIV infection or of reducing HIV burden. The term
"pharmaceutically effective amount" also refers to an amount
effective in treating, preventing or reducing the risk or
probability of developing acquired immunodeficiency syndrome
(AIDS), for delaying the apparition of AIDS, or reducing AIDS
symptoms. It is also to be understood herein that a
"pharmaceutically effective amount" may be construed as an amount
giving a desired therapeutic effect, either taken into a single or
multiple doses or in any dosage or route or taken alone or in
combination with other therapeutic agents. In the case of the
present invention, a "pharmaceutically effective amount" may be
understood as an amount having an inhibitory effect (partial or
complete) on HIV (HIV-1 and HIV-2 as well as related viruses (e.g.,
HTLV-I and HTLV-II, and simian immunodeficiency virus (SIV)))
infection cycle (e.g., inhibition of replication, reinfection,
maturation, budding etc.) and on any organism which rely on
aspartyl proteases for its life cycle. An inhibitory effect is to
be understood herein as an effect such as a reduction in the
capacity of an organism (e.g. HIV) to reproduce itself (replicate),
to re-infect surrounding cells, etc, or even a complete inhibition
(or elimination) of an organism.
[0309] The terms "HIV protease" and "HIV aspartyl protease" are
used interchangeably and includes, for example, the aspartyl
protease encoded by the human immunodeficiency virus type 1 or
2.
[0310] The term "prophylactically effective amount" refers to an
amount effective in preventing or reducing the risk or probability
of HIV infection in a patient. As used herein, the term "patient"
or "individual" refers to a mammal, including for example, a
human.
[0311] The terms "pharmaceutically acceptable carrier",
"pharmaceutically acceptable adjuvant" and "physiologically
acceptable vehicle" refer to a non-toxic carrier or adjuvant that
may be administered to a patient, together with one or more
compounds of the present invention, and which does not destroy the
pharmacological activity thereof.
[0312] The term "precursor" refers to a compound, such as a
Lysine-based compound which is able to be converted into an active
ingredient in vitro or in vivo. For example, the compound PL-461 is
a precursor of compound PL-100 as when administered to an
individual, PL-461 is converted into PL-100 in vivo (e.g., under
physiological conditions).
[0313] The term "derivative" refers to a compound which has been
chemically synthesized from an original compound. For example, when
considering the chemical synthesis of PL-461, PL-461 is a
derivative of PL-100.
[0314] The term "consisting essentially of" means that the
pharmaceutical composition includes the specified materials and may
include other material that does not materially affect the basic
characteristics of the pharmaceutical composition.
[0315] Pharmaceutically acceptable derivatives of the compounds of
formula I (such as compounds of formulae I, II, IIa, IIb, IIc, IIA
and IIA') and where applicable pharmaceutically acceptable salts
thereof such as, for example, ammonium salts are described herein.
A "pharmaceutically acceptable derivative" means any
pharmaceutically acceptable salt, ester, or salt of such ester, of
a compound of this invention or any other compound which, upon
administration to a recipient (a mammal), is capable of providing
(directly or indirectly) a an active compound or an antivirally
active metabolite or residue thereof.
[0316] It is to be understood herein that a "straight alkyl group
of 1 to 6 carbon atoms" includes for example, methyl, ethyl,
propyl, butyl, pentyl, hexyl.
[0317] It is to be understood herein that a "branched alkyl group
of 3 to 6 carbon atoms" includes for example, without limitation,
iso-butyl, tert-butyl, 2-pentyl, 3-pentyl, etc.
[0318] It is to be understood herein, that a "cycloalkyl group
having 3 to 6 carbon" includes for example, without limitation,
cyclopropyl, cyclobutyl, cyclopentyl, cyclocyclohexyl (i.e.,
C.sub.6H.sub.11).
[0319] Salts derived from appropriate bases include alkali metal
(e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium
and N--(C.sub.1-4 alkyl).sub.4.sup.+ salts.
[0320] The compounds described herein contain one or more
asymmetric carbon atoms and thus may occur as racemates and racemic
mixtures, single enantiomer, diastereomeric mixtures and individual
diastereoisomers. All such isomeric forms of these compounds are
expressly included in the present invention. Each stereogenic
carbon may be of the R or S configuration.
[0321] Pharmaceutically acceptable salts of the compounds described
herein include those derived from pharmaceutically acceptable
inorganic and organic acids and bases. Examples of such acid salts
include: acetate, adipate, alginate, aspartate, benzoate,
benzenesulfonate, bisulfate, butyrate, citrate, camphorate,
camphorsulfonate, cyclopentanepropionate, digluconate,
dodecylhydrogensulfate, dodecylsulfate, ethanesulfonate, formate,
fumarate, glucoheptanoate, glycerophosphate, glycollate,
hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,
hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate,
methanesulfonate, 2-naphthylsulfonate, nicotinate, nitrate,
oxalate, pamoate, pectinate, perchlorate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate,
and undecanoate.
[0322] Compounds which are encompassed by the present invention
also envisions the quaternization of any basic nitrogen containing
groups of the compounds disclosed herein. The basic nitrogen may be
quaternized with any agents known to those of ordinary skill in the
art including, for example, lower alkyl halides, such as methyl,
ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl
sulfates including dimethyl, diethyl, dibutyl and diamyl sulfates;
long chain halides such as decyl, lauryl, myristyl and stearyl
chlorides, bromides and iodides, and aralkyl halides including
benzyl and phenethyl bromides. Water or oil-soluble or dispersible
products may be obtained by such quaternization.
[0323] It is to be understood herein, that if a "range" or "group
of substances" is mentioned with respect to a particular
characteristic (e.g., temperature, concentration, time and the
like) of the present invention, the present invention relates to
and explicitly incorporates herein each and every specific member
and combination of sub-ranges or sub-groups therein whatsoever.
Thus, any specified range or group is to be understood as a
shorthand way of referring to each and every member of a range or
group individually as well as each and every possible sub-ranges or
sub-groups encompassed therein; and similarly with respect to any
sub-ranges or sub-groups therein. Thus, for example, [0324] with
respect to the number of carbon atoms, the mention of the range of
1 to 6 carbon atoms is to be understood herein as incorporating
each and every individual number of carbon atoms as well as
sub-ranges such as, for example, 1 carbon atoms, 3 carbon atoms, 4
to 6 carbon atoms, etc. [0325] with respect to reaction time, a
time of 1 minute or more is to be understood as specifically
incorporating herein each and every individual time, as well as
sub-range, above 1 minute, such as for example 1 minute, 3 to 15
minutes, 1 minute to 20 hours, 1 to 3 hours, 16 hours, 3 hours to
20 hours etc.; [0326] and similarly with respect to other
parameters such as concentrations, elements, etc. . . .
[0327] It is in particular to be understood herein that the
compound formulae each include each and every individual compound
described thereby as well as each and every possible class or
sub-group or sub-class of compounds whether such class or sub-class
is defined as positively including particular compounds, as
excluding particular compounds or a combination thereof; for
example an exclusionary definition for the formula (e.g. I) may
read as follows: "provided that when one of A and B is --COOH and
the other is H, --COOH may not occupy the 4' position".
[0328] It is also to be understood herein that "g" or "gm" is a
reference to the gram weight unit and "C", or ".degree. C." is a
reference to the Celsius temperature unit.
[0329] The compounds described herein may easily be prepared using
conventional techniques from readily available starting materials.
The detailed descriptions of these approaches are presented, for
example, in schemes 1 to 5 discussed below.
[0330] Scheme 1 illustrates a generic example for the preparation
of the phosphate monoester III derived from a primary alcohol (see
I), a compound of HIV protease inhibitors (see example 1 (step G
and H) in the experimental portion of this document for a specific
example of this synthesis).
[0331] Note:
[0332] a) R.sub.2 and R.sub.3 are as defined herein.
[0333] The synthesis of phosphate monoester III may use a HIV
aspartyl protease inhibitor (I, see U.S. Pat. No. 6,632,816) as the
starting material. The diethyl phosphotriester II was obtained in
good yield upon treatment with diethyl chlorophosphate and sodium
hydride in a mixture of tetrahydrofuran and triethylphosphate.
Then, addition of trimethysilyl bromide in dichloromethane (DCM)
gave compound III in good to excellent yields. ##STR95##
[0334] Scheme 1A represents another generic example for the
preparation of the phosphate monoester IIIA derived from a primary
alcohol (see IA), a compound of HIV protease inhibitors.
[0335] Note:
[0336] a) n, X, Y, R.sub.2, R.sub.3 and R.sub.6 are as defined
herein. ##STR96##
[0337] The synthesis of phosphate monoester IIIA is performed as
described for the preparation of III (scheme 1).
[0338] Scheme 2 illustrates a generic example for the preparation
of the phosphate monoester III, a compound of HIV protease
inhibitors, with a different approach starting from
(3S)-3-isobutylamino-azepan-2-one (IV).
[0339] Note:
[0340] a) R.sub.2 and R.sub.3 are as defined herein.
[0341] As shown in scheme 2, the phosphate monoester derivative III
was obtained from (3S)-3-isobutylamino-azepan-2-one (IV) in a
seven-step reaction sequence. Initially,
(2S)-3-isobutylamino-azepan-2-one (IV) was sulfonated with
4-acetamidobenzenesulfonyl chloride in the presence of
triethylamine in dichloromethane to give compound V in excellent
yields. The derivative VI was obtained quantitatively upon
treatment of V with di-tert-butyl pyrocarbonate and DMAP in
acetonitrile. The reductive ring opening with sodium borohydride in
ethanol lead to key intermediates VI in good yield. The diethyl
phosphotriester VIII was obtained in good yield upon treatment with
diethyl chlorophosphate and sodium hydride in a mixture of
tetrahydrofuran and triethylphosphate. The Boc protective groups
were removed upon treatment with HCl in ethanol to give compound IX
quantitatively (T. W. Greene and P. G. M. Wuts, Protective groups
in Organic Synthesis, 3.sup.rd Edition, John Wiley & Sons, Inc.
1999). Then, coupling of the free amino group present on
intermediate IX with a variety of synthetic amino acid in the
presence of 1-hydroxybenzotriazole (HOBt) and
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride
(EDAC) led to derivative II in good to excellent yields. Finally,
addition of trimethysilyl bromide in dichloromethane (DCM) gave
compound III in good to excellent yields. ##STR97##
[0342] Scheme 3 presents the transformation of a diphenylmethyl
derivative;
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-hydroxy-hexylc-
arbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl ester (PL-100)
into its fluorinated phosphate monoester sodium salt analog XI.
This reaction sequence may be used to produce any other similar
compounds (compounds) made of unsubstituted (or substituted)
diphenylmethyl, 1-naphthyl, 2-naphthyl, biphenyl and 9-anthryl
groups described herein.
[0343] Thus, the treatment of PL-100 with Selectfluor.TM. in
acetonitrile gave derivative X in 38% yield. The introduction of
the phosphate monoester group was performed as described previously
in scheme 1 and 2. First, the diethyl phosphotriester intermediate
was obtained in good yield upon treatment with diethyl
chlorophosphate and sodium hydride in a mixture of tetrahydrofuran
and triethylphosphate. Secondly, addition of trimethysilyl bromide
in dichloromethane (DCM) gave the phosphate monoester compound in
good to excellent yields. The final product XI was easily obtained
upon treatment of the phosphate monoester with a solution of sodium
hydroxide with good yields. ##STR98##
[0344] Scheme 4 illustrates a generic example for the
transformation of a phosphotriester II into its fluorinated analog
XIII in a two-step reaction sequence. This generic example
represents a second approach for the synthesis of fluorinated
compounds described herein. In this case, the fluorine atom is
added to the phosphotriester II instead of the primary alcohol
derivative of general formula or, more specifically, PL-100 as
shown on scheme 3. This alternate reaction sequence may be used to
produce any other similar compounds made of unsubstituted (or
substituted) diphenylmethyl, 1-naphthyl, 2-naphthyl, biphenyl and
9-anthryl groups described herein.
[0345] Note:
[0346] a) R.sub.2 and R.sub.3 are as defined herein.
[0347] Briefly, treatment of derivative II with Selectfluor.TM. in
acetonitrile gave derivative XII in good yields. Then, addition of
trimethysilyl bromide in dichloromethane (DCM) gave the phosphate
monoester compound XIII in good to excellent yields. If desired,
the final product XIII may be easily transformed into the phosphate
monoester sodium salt analog as described before in scheme 3.
##STR99##
[0348] Scheme 5 illustrates exemplary synthesis of various ester
compounds XVI described herein. The ester compounds are known to be
easily cleaved in vivo by esterase enzymes and, as a result, may
release the active ingredient. In this scheme R.sub.2 is set as a
diphenylmethyl group. However, this reaction sequence may be used
to produce any other similar compounds made of unsubstituted (or
substituted) diphenylmethyl, 1-naphthyl, 2-naphthyl, biphenyl and
9-anthryl groups described herein.
[0349] Note:
[0350] a) R.sub.1A represents the "residue" of the acid molecule
that is linked to the free primary alcohol group present on
intermediate XV and is as defined herein.
[0351] The compounds XVI are generally obtained in a three-step
reaction sequence in high yields. Esterification of
(1S)-{4-[(5-tert-butoxycarbonylamino-1-hydroxymethyl-pentyl)-isobutyl-sul-
famoyl]-phenyl}-carbamic acid tert-butyl ester (VII) with a variety
of acid in the presence of 1-hydroxybenzotriazole (HOBt) and
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride
(EDAC) led to the desired esters XIV in excellent yields. The
acetyl ester was obtained quantitatively using acetic anhydride in
the presence of N,N-dimethylaminopyridine (DMAP) in dichloromethane
(DCM). Cleavage of the Boc protective group was achieved
quantitatively upon treatment with trifluoroacetic acid (TFA) in
DCM. A second coupling with
(2S)-2-methoxycarbonylamino-3,3-diphenyl-propionic acid is
performed on the primary amino group of intermediate XV with HOBt
and EDAC to give the desired compounds XVI in good to excellent
yields. If necessary, catalytic hydrogenation of a
benzyloxycarbonyl group is performed using 10% palladium on carbon
to give the final compound XVII. ##STR100##
[0352] As it may be appreciated by the person skilled in the art,
the above synthetic schemes are not intended to be a comprehensive
list of all means by which the compound described in this
application may be synthesized but only represent exemplification
of synthesis methods among others. Further methods will be evident
to those of ordinary skill in the art.
[0353] The compounds described herein may be modified by appending
appropriate functionalities to enhance selective biological
properties. Such modifications may include those which increase
biological penetration into a given biological system (e.g., blood,
lymphatic system, central nervous system), increase oral
availability, increase solubility to allow administration by
injection, alter metabolism and alter rate of excretion.
[0354] As discussed above, the Lysine-based compounds may release
active ingredients which are excellent ligands for aspartyl
proteases, for example, HIV-1 protease. Accordingly, these
compounds are, by releasing the active ingredient, also capable of
targeting and inhibiting late stage events in the replication, i.e.
the processing of the viral polyproteins by HIV encoded protease.
Compounds described herein advantageously inhibit the ability of
the HIV-1 virus to infect immortalized human T cells over a period
of days, as determined by an assay measuring the amount of
extracellular p24 antigen; a specific marker of viral replication.
(see, Meek et al., Nature, 343, pp. 90-92 (1990)).
[0355] In addition to their use in the prophylaxis or treatment of
HIV or HTLV infection, the combination described herein may also be
used as inhibitory or interruptive agents for other viruses which
use aspartyl proteases, similar to HIV or HTLV aspartyl proteases,
in their life cycle. Such compounds inhibit the proteolytic
processing of viral polyprotein precursors by inhibiting aspartyl
protease. Because aspartyl protease is essential for the production
of mature virions, inhibition of that processing effectively blocks
the spread of virus by inhibiting the production and reproduction
of infectious virions, particularly from acutely and chronically
infected cells. The compounds described herein advantageously
inhibit aspartyl proteases, thus blocking the ability of aspartyl
proteases to catalyze the hydrolysis of peptide bonds.
[0356] The combination described herein may be employed in a
conventional manner for the treatment or prevention of HIV, HTLV,
and other viral infections, which involve aspartyl proteases for
their life (replication) cycle. Such methods of treatment, their
dosage levels and requirements may be selected by those of ordinary
skill in the art from available methods and techniques. For
example, combination of a compound as described herein and a CYP450
inhibitor may be associated with a pharmaceutically acceptable
adjuvant for administration to a virally infected patient in a
pharmaceutically acceptable manner and in an amount effective to
lessen the severity of the viral infection.
[0357] Alternatively, the combination described herein may be used
in vaccines and methods for protecting individuals against viral
infection over an extended period of time. The combination may be
employed in such vaccines either alone or together with other
compounds of this invention in a manner consistent with the
conventional utilization of protease inhibitors or protease
inhibitors derivatives in vaccines. For example, combination of a
compound as described herein with a CYP450 inhibitor may be
combined with pharmaceutically acceptable adjuvants, or delivery
systems conventionally employed in vaccines and administered in
prophylactically effective amounts to protect individuals over an
extended period of time against viral infections, such as HIV
infection. As such, the novel compounds of the present invention
(upon cleavage of a physiologically cleavable unit) may be
administered in combination with a CYP450 inhibitor as therapeutic
agents for treating or preventing viral infections, including HIV
infection, in a mammal and including AIDS treatment or
prevention.
[0358] The combination of compounds described herein with a CYP450
inhibitor may be administered to a healthy or HIV-infected patient
(before or after the onset of AIDS symptoms) either as a single
agent or in combination with other antiviral agents which interfere
with the replication cycle of HIV. By administering the compounds
of this invention with other antiviral agents which target
different events in the viral life cycle, the therapeutic effect of
these compounds is potentiated. For instance, the co-administered
antiviral agent may be one which targets early events in the viral
life cycle, such as attachment to the cell receptor and cell entry,
reverse transcription and viral DNA integration into cellular DNA.
Antiviral agents targeting such early life cycle events include
among others polysulfated polysaccharides, sT4 (soluble CD4) and
other compounds which block binding of virus to CD4 receptors on
CD4 bearing T-lymphocytes and other CD4(+) cells, or inhibit fusion
of the viral envelope with the cytoplasmic membrane, and didanosine
(ddI), zalcitabine (ddC), stavudine (d4T), zidovudine (AZT) and
lamivudine (3TC) which inhibit reverse transcription. For example
another protease inhibitor may be used with compounds of the
present invention. Other anti-retroviral and antiviral drugs may
also be co-administered with the compounds of this invention to
provide therapeutic treatment for substantially reducing or
eliminating viral infectivity and the symptoms associated
therewith. Examples of other antiviral agents include ganciclovir,
dideoxycytidine, trisodium phosphonoformate, eflomithine,
ribavirin, acyclovir, alpha interferon and trimenotrexate.
Additionally, other types of drugs may be used to potentiate the
effect of the compounds of this invention, such as viral uncoating
inhibitors, inhibitors of Tat or Rev trans-activating proteins,
antisense molecules or inhibitors of the viral integrase. These
compounds may also be co-administered with other inhibitors of HIV
aspartyl protease. Furthermore, it may be found useful to
administer compounds of the present invention with any other drug
(other anti-viral compounds, antibiotics, pain killer, etc.,).
[0359] Combination therapies according to this invention exert a
synergistic effect in inhibiting HIV replication because each
component agent of the combination acts on a different site of HIV
replication. The use of such combinations also advantageously
reduces the dosage of a given conventional anti-retroviral agent
that would be required for a desired therapeutic or prophylactic
effect as compared to when that agent is administered as a
monotherapy. These combinations may reduce or eliminate the side
effects of conventional single anti-retroviral agent therapies
while not interfering with the anti-retroviral activity of those
agents. These combinations reduce the potential of resistance to
single agent therapies, while minimizing any associated toxicity.
These combinations may also increase the efficacy of the
conventional agent without increasing the associated toxicity.
Combination therapies encompassed by the present invention include,
for example, the administration of a compound of this invention
with AZT, 3TC, ddI, ddC, d4T or other reverse transcriptase
inhibitors.
[0360] Alternatively, a combination of compounds described herein
with a CYP450 inhibitor may also be co-administered with other HIV
protease inhibitors such as Ro 31-8959 (Saquinavir; Roche),
L-735,524 (Indinavir; Merck), AG-1343 (Nelfinavir; Agouron),
ABT-378/r (Lopinavir; Abbott), and VX-478 (Amprenavir; Glaxo) to
increase the effect of therapy or prophylaxis against various viral
mutants or members of other HIV quasi species.
[0361] Administration of compounds of the present invention may be
performed, for example, as single agents or in combination with
retroviral reverse transcriptase inhibitors, or other HIV aspartyl
protease inhibitors. Co-administration of the compounds of this
invention with retroviral reverse transcriptase inhibitors or HIV
aspartyl protease inhibitors may exert a substantial synergistic
effect, thereby preventing, substantially reducing, or completely
eliminating viral infectivity and its associated symptoms.
[0362] The compounds of the present invention may be administered
in such a manner or form which may allow cleavage of the R.sub.1
unit to release a protease inhibitor. The compounds of this
invention may also be administered, for example, in combination
with immunomodulators (e.g., bropirimine, anti-human alpha
interferon antibody, IL-2, GM-CSF, methionine enkephalin,
interferon alpha, diethyldithiocarbamate sodium, tumor necrosis
factor, naltrexone and rEPO) antibiotics (e.g., pentamidine
isethionate) or vaccines to prevent or combat infection and disease
associated with HIV infection, such as AIDS and ARC.
[0363] When the combination of compounds described herein with a
CYP450 inhibitor are administered in combination therapies with
other agents, they may be administered sequentially or concurrently
to the patient. Alternatively, pharmaceutical or prophylactic
compositions according to this invention may be comprised of a
combination of one or more compounds of this invention and another
therapeutic or prophylactic agent.
[0364] Although this invention focuses on the use of the
combination disclosed herein for preventing and treating HIV
infection, the combination of this invention may also be used as
inhibitory agents for other viruses that depend on similar aspartyl
proteases for obligatory events in their life cycle. These viruses
include, but are not limited to, retroviruses causing AIDS-like
diseases such as simian immunodeficiency viruses, HIV-2, HTLV-I and
HTLV-II. In addition, the combination of this invention may also be
used to inhibit other aspartyl proteases and, in particular, other
human aspartyl proteases including renin and aspartyl proteases
that process endothelin precursors.
[0365] Pharmaceutical compositions of this invention comprise any
of the compounds of the present invention, and pharmaceutically
acceptable salts thereof, with any pharmaceutically acceptable
carrier, adjuvant or vehicle. Pharmaceutically acceptable carriers,
adjuvants and vehicles that may be used in the pharmaceutical
compositions of this invention include, but are not limited to ion
exchangers, alumina, aluminum stearate, lecithin, serum proteins,
such as human serum albumin, buffer substances such as phosphates,
glycine, sorbic acid, potassium sorbate, partial glyceride mixtures
of saturated vegetable fatty acids, water, salts or electrolytes,
such as protamine sulfate, disodium hydrogen phosphate, potassium
hydrogen phosphate, sodium chloride, zinc salts, colloidal silica,
magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based
substances, polyethyleneglycol, sodium carboxymethylcellulose,
polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,
polyethylene glycol and wool fat.
[0366] The pharmaceutical compositions of this invention may be
administered orally, parenterally by inhalation spray, topically,
rectally, nasally, buccally, vaginally or via an implanted
reservoir. It is therefore understood herein that oral
administration or administration by injection are encompassed by
the present invention. For example, compounds of the present
invention, may, for example, be orally administered in an aqueous
solution. The pharmaceutical compositions of this invention may
contain any conventional non-toxic pharmaceutically acceptable
carriers, adjuvants or vehicles. The term "parenteral" as used
herein includes subcutaneous, intracutaneous, intravenous,
intramuscular, intra-articular, intrasynovial, intrasternal,
intrathecal, intralesional and intracranial injection or infusion
techniques.
[0367] The pharmaceutical compositions may be in the form of a
sterile injectable preparation, for example, as a sterile
injectable aqueous or oleaginous suspension. This suspension may be
formulated according to techniques known in the art using suitable
dispersing or wetting agents (such as, for example, Tween 80) and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a non-toxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are amino acid, water, Ringer's
solution and isotonic sodium chloride solution. In addition,
sterile, fixed oils are conventionally employed as a solvent or
suspending medium. For this purpose, any bland fixed oil may be
employed including synthetic mono- or diglycerides. Fatty acids,
such as oleic acid and its glyceride derivatives are useful in the
preparation of injectables, as are natural
pharmaceutically-acceptable oils, such as olive oil or castor oil,
especially in their polyoxyethylated versions. These oil solutions
or suspensions may also contain a long-chain alcohol diluent or
dispersant, such as Ph. Helv. or a similar alcohol.
[0368] The pharmaceutical compositions of this invention may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, and aqueous suspension and
solutions. In the case of tablets for oral use, carriers that are
commonly used include lactose and corn starch. Lubricating agents,
such as magnesium stearate, are also typically added. For oral
administration in a capsule form, useful diluents include lactose
and dried cornstarch. When aqueous suspensions are administered
orally, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening and/or flavoring
and/or coloring agents may be added.
[0369] The pharmaceutical compositions of this invention may also
be administered in the form of suppositories for rectal
administration. These compositions may be prepared by mixing a
compound of this invention with a suitable non-irritating excipient
which is solid at room temperature but liquid at the rectal
temperature and therefore will melt in the rectum to release the
active components. Such materials include, but are not limited to,
cocoa butter, beeswax, and polyethylene glycols.
[0370] Topical administration of the pharmaceutical compositions of
this invention is especially useful when the desired treatment
involves areas or organs readily accessible by topical application.
For application topically to the skin, the pharmaceutical
composition should be formulated with a suitable ointment
containing the active components suspended or dissolved in a
carrier. Carriers for topical administration of the compounds of
this invention include, but are not limited to, mineral oil, liquid
petroleum, white petroleum, propylene glycol, polyoxyethylene or
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutical compositions may be formulated
with a suitable lotion or cream containing the active compound
suspended or dissolved in a carrier. Suitable carriers include, but
are not limited to, mineral oil, sorbitan monostearate, polysorbate
60, cetyl esters wax cetearyl alcohol, 2-octyldodecanol, benzyl
alcohol and water. The pharmaceutical compositions of this
invention may also be topically applied to the lower intestinal
tract by rectal suppository formulation or in a suitable neat
formulation. Topically-transdermal patches are also included in
this invention.
[0371] The pharmaceutical compositions of this invention may be
administered by nasal aerosol or inhalation. Such compositions are
prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other solubilizing or dispersing agents known in the
art.
[0372] Dosage levels of between about 0.01 and about 150 mg/kg body
weight per day of for example, 0.01 and about 50 mg/kg body weight
per day or for example from between about 0.5 and about 30 mg/kg
body weight per day of the active ingredient compound are useful in
the prevention and treatment of viral infection, including HIV
infection. Typically, the pharmaceutical compositions of this
invention will be administered from about 1 to about 5 times per
day or alternatively, as a continuous infusion. Such administration
may be used as a chronic or acute therapy. The amount of active
ingredient that may be combined with the carrier materials to
produce a single dosage form will vary depending upon the patient
treated and the particular mode of administration. A typical
preparation will contain from about 5% to about 95% of a desired
compound (w/w) i.e., active ingredient or a precursor. For example,
such preparations may contain from about 20% to about 80% of a
desired compound.
[0373] Upon improvement of a patient's condition, a maintenance
dose of a compound, composition or combination of this invention
may be administered if necessary. Subsequently, the dosage or
frequency of administration, or both, may be reduced, as a function
of the symptoms, to a level at which the improved condition is
retained. When the symptoms have been alleviated to the desired
level, treatment should cease. Patients may, however, require
intermittent treatment on a long-term basis, upon any recurrence of
disease symptoms.
[0374] As the person skilled in the art will appreciate, lower or
higher doses than those recited above may be desired. Specific
dosage and treatment regimen for any particular patient may depend
upon a variety of factors, including the activity of the specific
compound employed, the age, body weight, general health status,
sex, diet, time of administration, rate of excretion, drug
combination, the severity and course of the infection, the
patient's disposition to the infection and the judgment of the
treating physician.
[0375] In the description herein, the following abbreviations are
used:
[0376] Abbreviation Meaning [0377] Ac Acetyl [0378] AcOH Acetic
acid [0379] APCl Atmospheric pressure chemical ionization [0380]
AIDS Acquired Immunodeficiency Syndrome [0381] APV Amprenavir
[0382] ATV Atazanavir [0383] AZT 3-Azido-3-deoxythymine
(Zidovudine) [0384] Boc Benzyloxycarbonyl [0385] t-Butyl tert-Butyl
[0386] CAM Cerium ammonium molybdate [0387] DCM Dichloromethane
[0388] DMAP N,N-dimethylaminopyridine [0389] DMSO Dimethylsulfoxide
[0390] DMF Dimethylformamide [0391] DNA Deoxyribonucleic acid
[0392] EDAC 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide
hydrochloride [0393] EtOAc Ethyl acetate [0394] EtOH Ethyl alcohol
[0395] g Gram [0396] h hour [0397] HIV-1, -2 Human immunodeficiency
virus type 1, type 2 [0398] HOBt 1-Hydroxybenzotriazole [0399] HPLC
High performance liquid chromatography [0400] HTLV-I, -II Human
T-cell lymphotropic virus type I, type II [0401] IL-2 Interleukin-2
[0402] IDV Indinavir [0403] Kg Kilogram [0404] L Liter [0405] LC-MS
Liquid chromatography-mass spectrometry [0406] LPV Lopinavir [0407]
M Molar [0408] MeOH Methyl alcohol [0409] mg Milligram [0410] mp
Melting point [0411] min Minute [0412] Moc Methoxycarbonyl [0413]
mol Mole [0414] mL Milliliter [0415] mmol Millimole [0416] NFV
Nelfinavir [0417] nm Nanometer [0418] nM Nanomolar [0419] po Orally
[0420] rEPO Recombinant erythropoietin [0421] RTV Ritonavir [0422]
SQV Saquinavir [0423] TLC Thin layer chromatography [0424] 3TC
2',3'-Dideoxy-3-thiacytidine [0425] TFA Trifluoroacetic acid [0426]
THF Tetrahydrofuran
BRIEF DESCRIPTION OF THE DRAWINGS
[0427] In drawings which illustrates exemplary embodiments of the
present invention;
[0428] FIG. 1 is a graph illustrating the fold-change (FC) and
median fold-change in IC.sub.50 vs the reference strain (NL-4.3)
observed for various protease inhibitors;
[0429] FIG. 2 is a schematic illustrating the advantages of the
Lysine-based compounds disclosed herein;
[0430] FIG. 3 is a schematic illustrating the cleavage of the
physiologically cleavable unit of a Lysine-based compound (e.g.,
PL-461) generating the active ingredient (e.g., PL-100);
[0431] FIG. 4 is a graph illustrating water solubility of PL-461 at
various pH;
[0432] FIG. 5 is a graph illustrating the plasma concentration of
the drug after administration of either a Lysine-based compound
(e.g., PL-461) or an active ingredient (e.g., PL-100) in rats;
[0433] FIG. 6 is a histogram illustrating the effect of CYP450
inhibitors on the metabolism of various HIV-1 protease inhibitors
in human liver microsomes;
[0434] FIG. 7 is a histogram illustrating the bioavailability of a
Lysine-based compound (e.g., PL-461) or of an active ingredient
(e.g., PL-100) in combination with various concentration of a
CYP450 inhibitor (e.g., ritonavir) in rats;
[0435] FIG. 8 is a graph illustrating the plasma concentration of
the drug after administration of a Lysine-based compound (e.g.,
PL-461) in combination with a CYP450 inhibitor (e.g., ritonavir) in
rats;
[0436] FIG. 9 is a histogram illustrating the bioavailability of
the drug following administration of various Lysine-based compound
(e.g., PL-461)/CYP450 inhibitor (CYP450) ratios in rats and;
[0437] FIG. 10 is a table illustrating the inhibition of CYP450
sub-families by the active ingredient PL-100.
EXAMPLES
[0438] This section describes the synthesis of lysine based
compounds able to release an HIV aspartyl protease inhibitors as
described herein. These examples are for the purpose of
illustration only and are not to be construed as limiting the scope
of the invention in any way. This section presents the detailed
synthesis of compounds no. 1 to 10 of this invention.
[0439] Exemplary synthesis schemes of the active ingredients have
been disclosed in U.S. Pat. No. 6,632,816 to Stranix et al.
[0440] Materials and Methods-Preparation of Compounds
[0441] Analytical thin layer chromatography (TLC) was carried out
with 0.25 mm silica gel E. Merck 60 F.sub.254 plates and eluted
with the indicated solvent systems. Preparative chromatography was
performed by flash chromatography, using silica gel 60 (EM Science)
with the indicated solvent systems and positive air pressure to
allow proper rate of elution. Detection of the compounds was
carried out by exposing eluted plates (analytical or preparative)
to iodine, UV light and/or treating analytical plates with a 2%
solution of p-anisaldehyde in ethanol containing 3% sulfuric acid
and 1% acetic acid followed by heating. Alternatively, analytical
plates may be treated with a 0.3% ninhydrin solution in ethanol
containing 3% acetic acid and/or a CAM solution made of 20 g
(NH.sub.4).sub.6Mo.sub.7O.sub.24 and 8.3 g Ce(SO.sub.4).sub.2
polyhydrate in water (750 mL) containing concentrated sulfuric acid
(90 mL).
[0442] Preparative HPLC were performed on a Gilson apparatus
equipped with a C18 column, a 215 liquid handler module and 25
mL/min capacity head pumps. The HPLC is operated with a Gilson
UniPoint System Software.
[0443] Semi-Preparative HPLC Conditions for Purification of Test
Compounds:
[0444] HPLC system: 2 Gilson #305-25 mL pumps, Gilson #215 liquid
handler for injection and collection and a Gilson #155 UV-Vis
absorbance detector, all controlled from a Gilson Unipoint V1.91
software
[0445] Column: Alltech (#96053) Hyperprep PEP, C-18, 100
.ANG..alpha., 8 .mu.m, 22.times.250 mm
[0446] Flow: 15 mL/min
[0447] Solvents: A: H.sub.2O; B: CH.sub.3CN
[0448] Gradient: 25% to 80% of B over 40 min
[0449] Detector: absorbance; .lamda.: 210 & 265 nm
[0450] The crude material dissolved in acetonitrile to a
concentration of around 50 to 80 mg/2 mL were injected in each run.
Fractions were collected in amounts of 9 mL pertaining absorbance
was detected at the UV detector.
[0451] Unless otherwise indicated, all starting materials were
purchased from a commercial source such as Aldrich Co. or Sigma
Co.
[0452] Melting points (mp) were determined on a Buchi 530 melting
point apparatus in capillary tubes and were uncorrected.
[0453] Mass spectra were recorded on a Hewlett Packard LC/MSD 1100
system using APCl or electrospray sources either in negative mode
or positive mode.
[0454] Nuclear magnetic resonance (NMR) spectra were recorded on a
Bruker AMX-II-500 equipped with a reversed or QNP probe. Samples
were dissolved in deuterochloroform (CDCl.sub.3), deuteroacetone
(acetone-d.sub.6), deuteromethanol (CD.sub.3OD) or
deuterodimethylsulfoxide (DMSO-d.sub.6) for data acquisition using
tetramethylsilane as internal standard. Chemical shifts 0 are
expressed in parts per million (ppm), the coupling constants (J)
are expressed in hertz (Hz) whereas multiplicities are denoted as s
for singlet, d for doublet, 2d for two doublets, dd for doublet of
doublets, t for triplet, q for quartet, quint. for quintet, m for
multiplet, and br s for broad singlet.
DETAILED DESCRIPTION OF THE INVENTION
Examples
Specific Examples for the Preparation of Derivatives of General
Formula I
[0455] The preparation of PL-100, PL-337 and other compounds of
this class is presented in U.S. Pat. No. 6,632,816 to Stranix et
al.
[0456] The following compounds were prepared from L-lysine
derivatives using the procedures summarized in schemes 1, 1A, 2, 3,
4 and 5 of this invention.
Example 1
Preparation of
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-phosphonooxy-h-
exylcarbamoyl}-2,2-diphenyl-ethyl)-carbamic Acid Methyl Ester
(PL-461)
[0457] The preparation of the title compound is based on schemes 1
and 2 of this invention.
[0458] Step A. Preparation of (3S)-3-isobutylamino-azepan-2-one
(IV)
[0459] L-.alpha.-amino-caprolactam (22.0 g) was dissolved in cold
dichloroethane (DCM, 200 mL). isobutyraldehyde (12.6 g) was added
slowly and stirred until the heat evolved was dissipated (water
forms at the surface). The cold solution was added to 46.5 g of
powdered NaBH(OAc).sub.3 in DCM (0.5 L). AcOH (70 mL) was added to
the solution. The slightly turbid mixture was stirred at 20.degree.
C. for 4 h. A 500 mL solution of 2M NaOH was added slowly to the
turbid mixture and the pH adjust to 11 using a concentrated NaOH
solution, and then the mixture stirred for a further 20 min. After
extraction, the DCM layer was dried with MgSO.sub.4, filtered and
evaporated. The oil thus obtained crystallizes slowly on standing
(27.8 g, 85%) and was used without further purification in the next
step.
[0460] .sup.1H NMR (CDCl.sub.3): .delta. 0.93 (d, J=6.5, 3H), 0.97
(d, J=6.5, 3H), 1.39 (t, J=9.8, 1H), 1.47 (m, 1H), 1.78-1.65 (m,
2H), 2.00-1.93 (m, 2H), 2.32-2.2 (m, 2H), 2.38 (t, J=9.7, 1H), 3.16
(m, 3H), 6.62 (s, 1H(NH)). mp 52-54.degree. C. (hexanes).
[0461] A small sample was converted to the S-methyl benzyl urea by
adding the solid to a solution of S-methyl benzyl isocyanate in
MeCN. NMR gives 98% ee
[0462] Step B. Preparation of
N.alpha.-isobutyl-N.alpha.-(4-acetamidobenzenesulfonyl)-L-.alpha.-amino-c-
aprolactam (V)
[0463] N.alpha.-isobutyl-L-.alpha.-amino-caprolactam (IV) (4.1 g
free base) was dissolved in DCM (200 mL) and treated with 4.0 g
triethylamine, followed by 4-acetamidobenzenesulfonyl chloride (5.2
g). A 0.1 g portion of dimethylaminopyridine was added and the
mixture was stirred 5 h. The resulting thick slurry was poured into
500 mL 0.5 M HCl and shaken vigorously. The solid in the biphasic
solution was filtered out and washed with cold acetone to give 7.3
g (87%) of clean product.
[0464] .sup.1H NMR (DMSO-d.sub.6): 0.93 (d, J=6.0, 3H), 0.96 (d,
J=6.0, 3H), 1.39 (t, J=12.0, 1H), 1.85-1.65 (m, 3H), 2.08-2.18 (m
and s, 6H), 2.90-2.97 (m, 1H), 3.00-3.06 (m, 2H), 3.35 (dd, J=14.2,
8.5, 1H), 4.65 (d, J=8.7, 1H), 6.3 (s, 1H), 7.42 (d, J=8.8, 2H),
7.6 (d, J=8.8, 2H). mp 230-233.degree. C. (EtOH).
[0465] Step C. Preparation of
(3S)-3-{([4-(acetyl-tert-butoxycarbonyl-amino)-benzenesulfonyl]-isobutyl--
amino}-2-oxo-azepane-1-carboxylic acid tert-butyl ester (Boc
activation) (VI)
[0466] 4.2 g of
N.alpha.-isobutyl-N.alpha.-(4-acetamidobenzenesulfonyl)-L-.alpha.-amino-c-
aprolactam (V) was suspended in 30 mL MeCN and briefly sonicated to
break up any large chunks. To this white suspension was added 6.7 g
(3 eq.) of di-tert-butyl pyrocarbonate in 10 mL MeCN. The
suspension was stirred with a magnetic bar and a 120 mg portion of
DMAP was added. The solution becomes a clear light yellow after a
few minutes. TLC (EtOAc) reveals 1 product Rf 0.9 (starting
material Rf at 0.4). The solution is poured in distilled water 20
mL and extracted with ether, dried with Na.sub.2SO.sub.4 and
evaporated yielding 6.90 g. A sample was recrystallized from
hexanes.
[0467] .sup.1H NMR (DMSO-d.sub.6): 0.68 (d, J=6.0, 3H), 0.85 (d,
J=6.0, 3H), 1.39 (s, 10H), 1.47 (s, 9H), 1.85-1.65 (m, 3H), 2.15
(s, 3H), 2.80 (q, J=4, 1H), 3.10-3.36 (m, 2H), 4.01 (d, J=8.0, 1H),
4.85 (d, J=8.7, 1H), 7.32 (d, J=8.8, 2H), 7.87 (d, J=8.8, 2H). mp
123-124.degree. C.
[0468] Step D. Preparation of
(1S)-4-amino-N-(5-amino-1-hydroxymethyl-pentyl)-N-isobutyl-benzenesulfona-
mide (VII-deprotected) (Reductive Ring Opening and
Deprotection)
[0469] A 3.0 g portion of
(3S)-3-{[4-(acetyl-tert-butoxycarbonyl-amino)-benzenesulfonyl]-isobutyl-a-
mino}-2-oxo-azepane-1-carboxylic acid tert-butyl ester (VI, step C)
is dissolved in 40 mL EtOH followed by 750 mg NaBH.sub.4. Brief
heating with a heat gun gives a clear solution. TLC reveals one
streaky spot after 20 min (EtOAc). The solution is concentrated to
a paste, poured in 40 mL 1N NaOH and extracted with ethyl acetate,
the organic phase dried with NaSO.sub.4 and evaporated to give 2.8
g of product intermediate (VII);
(1S)-{4-[(5-tert-butoxycarbonylamino-1-hydroxymethyl-pentyl)-isobutyl-sul-
famoyl]-phenyl}-carbamic acid tert-butyl ester (VII).
[0470] The above product intermediate is dissolved in 5 mL EtOH and
5 mL 12 N HCl is added. Vigorous gas evolution is observed for a
few minutes. After 2 h the solution is evaporated and rendered
basic with concentrated KOH and extracted with EtOAc yielding 1.75
g of a white powder.
[0471] .sup.1H NMR (DMSO-d.sub.6): 0.82 (m, 6H), 0.97-1.12 (m, 2H),
1.15-1.30 (m, 3H), 1.57 (m, 1H), 1.84 (m, 1H), 2.40 (t, J=7.8, 2H),
2.75 (m, 1H), 2.85 (m, 1H), 3.21 (m, 1H), 3.44 (d, J=6.4, 2H), 5.92
(br s, 2H), 6.59 (d, J=8.0, 2H), 7.39 (d, J=8.0, 2H).
[0472] Step E. Preparation
(2S)-2-methoxycarbonylamino-3,3-diphenyl-propionic acid
[0473] To a solution of L-diphenylalanine (241 mg, 1.0 mmol)
(Peptech Corp.) in 5 mL 1N NaOH and 0.5 mL saturated
Na.sub.2CO.sub.3 (resulting solution at pH 10) was added
methoxycarbonyloxysuccinimide (carbonic acid
2,5-dioxo-pyrrolidin-1-yl ester methyl ester) (180 mg, 1.1 mmol)
dissolved in 5 mL. Afterwards, the reaction mixture was stirred at
room temperature for 2 h. The alkaline solution was extracted once
with ether (10 mL) and the aqueous phase was acidified with 1N HCl.
This was extracted twice with 20 mL EtOAc, and the combined organic
phases were washed with 50 mL 1N HCl. The organic phase was dried
over Na.sub.2SO.sub.4 filtered and evaporated to an oil, which
solidifies to yields 250 mg (83%) of the desired material. This
derivative was used as such in the next step.
[0474] Step F. Preparation of
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-hydroxy-hexylc-
arbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl ester
(PL-100)
[0475] The title compound was prepared from
(1S)-4-amino-N-(5-amino-1-hydroxymethyl-pentyl)-N-isobutyl-benzenesulfona-
mide (VII-deprotected) (step D) and
(2S)-2-methoxycarbonylamino-3,3-diphenyl-propionic acid (step E)
using the coupling procedure with HOBt and EDAC described in
example 3 (step D). The final product was obtained in 67% yield
(121 mg).
[0476] LC-MS: 625.3 (M+H).sup.+, 95% pure
[0477] .sup.1H NMR (CD.sub.3OD): .delta. 0.71-0.85 (m, 2H), 0.88
(d, J=6.3, 5H), 0.91-0.96 (m, 2H), 1.29-1.34 (m, 1H), 1.41-1.52 (m,
1H) 1.82-1.92 (m, 1H), 2.61-2.68 (m, 1H), 2.81-2.85 (m, 2H),
2.94-3.05 (m, 2H), 3.38-3.40 (t, J=5.0, 1H), 3.50-3.51 (m, 1H),
3.52 (s, 3H), 4.28 (d, J=11.0 1H), 4.87 (d, J=11.0, 1H), 6.69 (d,
J=8.0, 2H), 7.15-718 (m, 2H), 7.20-7.31 (m, 6H), 7.33 (d, J=7.9,
2H), 7.47 (d, J=7.5, 1H).
[0478] .sup.13C NMR (CD.sub.3OD): .delta. 20.0, 20.1, 23.3, 25.4,
28.1, 28.5, 28.9, 38.1, 40.0, 51.2, 51.6, 53.1, 57.2, 57.4, 59.5,
61.9, 62.4, 112.6, 125.7, 126.2, 126.3, 127.9, 128.1, 128.15,
128.2, 128.4, 128.7, 141.3, 141.9, 152.4, 155.9, 169.9, 172.5.
[0479] Step G. Preparation of
(1S,5S)-{1-[5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(diethoxy-phos-
phoryloxy)-hexylcarbamoyl]-2,2-diphenyl-ethyl}-carbamic acid methyl
ester
[0480] The PL-100 compound (product of step F, 203 mg, 0.325 mmol)
was dissolved in dry tetrahydrofuran (3 mL) and 0.2 mL
triethylphosphate under N.sub.2 atmosphere. The mixture was stirred
at this temperature for 15 min, followed by the addition of diethyl
chlorophosphate (0.061 mL, 0.423 mmol). Sodium hydride (60% in
mineral oil) (17 mg, 0.423 mmol) was added at 0.degree. C. The
solution was stirred for 1 h at 0.degree. C. and 12 h at room
temperature. 20 mL of Amberlite XAD-2 was added to the solution and
the beads were thoroughly mixed with the solvent. To the mixture
was added ice water 2 mL, and the THF evaporated off. The beads
were then washed with distilled water 6 times 100 mL then extracted
three times with ethyl acetate (30 mL). The combined phase was
evaporated and the residue was dried under high vacuum. The crude
product was purified by flash chromatography using ethyl
acetate/hexane (8/2), then EtOAc 100% as eluent. The yield of this
reaction is 152 mg 61%.
[0481] LC-MS: 761.2 (M+H).sup.+, 90% pure
[0482] .sup.1H NMR (CD.sub.3OD): .delta. 0.68-0.75 (m, 1H),
0.75-0.84 (m, 1H), 0.84-1.10 (m, 9H), 1.21-1.50 (m, 8H), 1.88 (m,
1H), 2.58-2.71 (m, 1H), 2.80-2.89 (m, 1H), 2.89-3.08 (m, 2H),
3.49-3.60 (s, 3H), 3.65-3.74 (m, 1H), 3.85-3.95 (m, 1H), 3.97-4.02
(m, 1H), 4.07-4.21 (m, 4H), 4.29 (d, J=10.8, 1H), 6.71 (d, J=8.0,
2H), 7.10-7.20 (m, 2H), 7.20-7.32 (m, 5H), 7.32-7.45 (m, 3H), 7.50
(d, J=7.5, 2H), 7.86 (br s, 1H).
[0483] .sup.31P NMR (CD.sub.3OD): .delta. 1.62
[0484] Step H. Preparation of
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-phosphonooxy-h-
exylcarbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl ester
(PL-461)
[0485] The product of step G prepared above (152 mg) was dissolved
in anhydrous dichloromethane (3.0 mL). Trimethylsilyl bromide (0.5
mL) was added at 0.degree. C. The mixture was stirred during 1 h at
this temperature and overnight at room temperature. The solvent was
evaporated and 0.2 mL water was added to the residue. 3 mL EtOH was
added mixed and evaporated. This step was repeated three times and
the residue dried in vacuo. Yields 98 mg 70% of the title
derivatives of this first example.
[0486] LC-MS: 705.2 (M+H).sup.+, 95% pure
[0487] .sup.1H NMR (CD.sub.3OD): .delta. 0.65-0.73 (m, 1H),
0.75-0.83 (m, 1H), 0.89 (d, J=5.6, 8H), 1.27-1.38, (m, 1H),
1.42-4.55 (m, 1H), 1.82-1.94 (m, 1H), 2.57-2.68 (m, 1H), 2.78-2.90
(m, 1H), 2.91-3.09 (m, 2H), 3.54 (s, 3H), 3.60-3.72 (m, 1H),
3.87-4.05 (m, 1H), 4.00 (m, 1H), 4.29 (d, J=11.3, 1H), 4.90 (d,
J=11.4, 1H), 6.73 (d, J=8.0, 2H), 7.13-7.22 (m, 2H), 7.22-7.33 (m,
6H), 7.33-7.45 (m, 2H), 7.51 (d, J=7.5, 2H).
[0488] .sup.31P NMR (CD.sub.3OD): .delta. 2.80
Example 2
Preparation of
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-phosphonooxy-h-
exylcarbamoyl}-2,2-diphenyl-ethyl)-carbamic Acid Methyl Ester
Sodium Salt (PL-462)
[0489] 70.7 mg of the final product of example 1 is added to 1 mL
0.1 N NaOH and diluted with 1 mL of distilled water. The Solution
is then frozen and lyophilized. Yields 67.2 mg (92%) of the desired
material with 95% purity.
[0490] .sup.1H NMR (CD.sub.3OD): .delta. 0.72-0.83 (m, 1H), 0.90
(d, J=5.8, 9H), 1.26-1.38 (m, 1H), 1.53-1.65 (m, 1H), 1.88-2.00 (m,
1H), 2.60-2.70 (m, 1H), 2.79-2.89 (m, 1H), 2.98-3.00 (m, 1H),
3.00-3.08 (m, 1H), 3.54 (s, 3H), 3.58-3.71 (m, 1H), 3.72-3.83 (m,
1H), 3.84-3.95 (m, 1H), 4.28 (d, J=11.1, 1H), 4.91 (d, J=11.0, 1H),
6.70 (d, J=7.6, 2H), 7.12-7.22 (m, 2H), 7.22-7.32 (m, 6H),
7.33-7.40 (m, 2H), 7.50 (d, J=7.7, 2H).
[0491] .sup.31P NMR (CD.sub.3OD): .delta. 3.13
Example 3
Preparation of
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-phosphonooxy-h-
exylcarbamoyl}-2-naphthalen-2-yl-ethyl)-carbamic Acid Methyl Ester
(PL-507)
[0492] The preparation of the title compound is based on scheme 2
of this invention.
[0493] Step A. Preparation of
(1S)-(4-{[5-tert-butoxycarbonylamino-1-(diethoxyphosphoryloxymethyl)-pent-
yl]-isobutyl-sulfamoyl)phenyl)-carbamic acid tert-butyl ester
(VII)
[0494] 2.00 g (3.7 mmol)
(1S)-(4-[(5-tert-butoxycarbonylamino-1-hydroxymethyl-pentyl)-isobutyl-sul-
famoyl]-phenyl}-carbamic acid tert-butyl ester (VII) (example 1,
step D) is dissolved in 0.63 mL triethylphosphate and 10 mL THF at
0.degree. C. under inert argon atmosphere. 0.63 mL (4.44 mmol)
diethylchlorophosphate is added and then 0.25 g (6.2 mmol), NaH 60%
in oil is added in portionwise. The mixture is allowed to warm to
room temperature and left to stir for 2 h (LC-MS showed completion
after 1 h). To the solution is added 20 mL of Amberlite XAD-2 resin
and the slurry thoroughly mixed and added to 200 mL ice water.
After stirring for 15 min. the resin suspension is filtered and the
resin washed several times with distilled water (500 mL). The
desired product is desorbed from the resin with acetone (5.times.50
mL), EtOAc (5.times.50 mL), the organic phase is then dried over
Na.sub.2SO.sub.4. After evaporation of the solvent 2.66 g (89%) of
clear oil is obtained. The crude product contains a fraction with
two diethyl phosphates and is used as is in the next step.
[0495] .sup.1H NMR (CD.sub.3OD): .delta. 0.91 (d, J=6.3, 6H),
1.11-1.21 (m, 2H), 1.33 (t, J=6.9, 10H), 1.43 (s, 9H), 1.53 (s,
10H), 1.90-1.97 (m, 1H), 2.88-2.96 (m, 3H), 2.96-3.04 (m, 1H),
3.81-3.90 (m, 1H), 3.91-3.99 (m, 1H), 4.01-4.14 (m, 4H), 7.61 (d,
J=8.3, 2H), 7.72 (d, J=8.4, 2H).
[0496] .sup.31P NMR (CD.sub.3OD): .delta. 1.59
[0497] Step B. Preparation of (2S)-phosphoric acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
diethyl ester (IX)
[0498] The crude product obtained in the previous step (VIII, 2.66
g) is dissolved in 12 mL EtOH. 4 mL of HCl.sub.conc, is added and
heated briefly to 70.degree. C. then left at room temperature for 3
h. The solvent is evacuated and the residue triturated with 50 mL
ether. The thick residue is then dissolved in 3 mL ice water and
the pH adjusted to 12 with 50% NaOH. The thick slurry obtained is
extracted with EtOAc (3.times.50 mL) and the organic phase dried
over Na.sub.2SO.sub.4. After filtration of the drying agent the
organic phase is evacuated to yield 1.84 g (98%) of the desired
product (IX).
[0499] LC-MS: 480.2 (M+H).sup.+, 95% pure.
[0500] .sup.1H NMR (CD.sub.3OD): .delta. 0.91 (s, 6H), 1.11-1.26
(m, 3H), 1.28-1.43 (m, 8H), 1.45-1.51 (m, 1H), 1.52-1.61 (m, 1H),
1.89-1.96 (m, 1H), 2.56 (t, J=6.7, 2H), 2.85-2.91 (m, 1H),
2.98-3.11 (m, 1H), 3.79-3.99 (m, 1H), 3.94 (d, J=5.3, 1H),
4.09-4.11 (m, 4H), 6.69 (d, J=7.9, 2H), 7.50 (d, J=7.9, 2H).
[0501] .sup.31P NMR (CD.sub.3OD): .delta.1.61
[0502] Step C. Preparation of
(2S)-2-methoxycarbonylamino-3-naphthalen-2-yl-propionic acid (or
L-Moc-2-naphthylalanine)
[0503] To a solution of L-2-naphthylalanine (215 mg, 1 mmol)
(Peptech Corp.) in 5 mL 1N NaOH and 0.5 mL saturated
Na.sub.2CO.sub.3 (resulting solution at pH 10) was added
methoxycarbonyloxysuccinimide (187 mg, 1.1 mmol) dissolved in 5 mL.
Afterwards, the reaction mixture was stirred at room temperature
for 2 h. The alkaline solution was extracted once with ether (10
mL) and the aqueous phase was acidified with 1N HCl. This was
extracted twice with 20 mL EtOAc, and the combined organic phases
were washed with 50 mL 1N HCl. The organic phase was dried over
Na.sub.2SO.sub.4, filtered and evaporated to an oil, which
solidifies to yields 200 mg (73%) of the desired material. This
intermediate (referred as the N-substituted amino acid) was used
without further purification in the next step.
[0504] Step D. Preparation of
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-phosphonooxy-h-
exylcarbamoyl}-2-naphthalen-2-yl-ethyl)-carbamic acid methyl ester
(PL-507)
[0505] 100 mg L-Moc-2-naphthylalanine (step C) was activated with
100 mg EDAC and 57 mg HOBt in 1.5 mL DMF for 30 minutes. Then, 100
mg of phosphoric acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
diethyl ester (step B) was added and left to stir at room
temperature for 1 h. 40 mL of 1M K.sub.2CO.sub.3 was added to the
DMF solution and left for 10 min. 50 mL of EtOAc was then added and
the mixture was then agitated vigorously. Separation of the EtOAc
phase was effected, followed by extraction with 5% citric acid (50
mL) once, then water (50 mL) 3 times and finally brine. The organic
phase was the separated and evaporated. The residue was taken up in
50 mL DCM and re-evaporated. The residue was again taken up in 50
mL DCM and 0.5 mL of TMSBr was added. The solution was left
overnight (16 h). The DCM was evacuated and a solution of ice cold
MeOH: Water 1:1 was added, stirred briefly and evacuated. The
residue was triturated with ether then dissolved in 1N NaOH. The
clear solution was extracted with ether and the aqueous phase
acidified with 6N HCl. The white precipitated was then collected by
filtration and dried in vacuo overnight. Yields 88 mg of the title
compound.
[0506] LC-MS: 679.8 (M+H).sup.+, 95% pure.
[0507] .sup.1H NMR (CD.sub.3OD): .delta. 0.89-0.98 (m, 8H), 1.15
(m, 2H), 1.35 (m, 1H), 1.45 (m, 1H), 1.88 (m, 1H), 2.84 (m, 2H),
2.98 (m, 1H), 3.01 (m, 2H), 3.24 (m, 1H), 3.56 (s, 3H), 3.60 (m,
1H), 3.81 (m, 1H), 3.99 (m, 1H), 4.39 (t, J=6.8, 1H), 6.91 (d,
J=8.0, 2H), 7.34 (d, J=8.0, 1H), 7.45 (m, 2H), 7.58 (d, J=8.0, 2H),
7.66 (s, 1H), 7.70-7.82 (m, 3H).
[0508] .sup.31P NMR (CD.sub.3OD): .delta. 2.56
Example 4
Preparation of (2S,2S)phosphoric acid
mono-(2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-{2-[(morpholine-4-ca-
rbonyl)-amino]-3-naphthalen-1-yl-propionylamino}-hexyl)ester
(PL-498)
[0509] Step A. Preparation of
(2S)-2-[(morpholine-4-carbonyl)-amino]-3-naphthalen-1-yl-propionic
acid
[0510] To a solution of L-1-naphthylalanine (215 mg, 1 mmol)
(Peptech Corp.) in 5 mL 1N NaOH and 0.5 mL saturated
Na.sub.2CO.sub.3 (resulting solution at pH 10) was added
morpholine-4-carbonyl chloride (150 mg, 1.0 mmol) dissolved in 5
mL. Afterwards, the reaction mixture was stirred at room
temperature for 2 h. The alkaline solution was extracted once with
ether (10 mL) and the aqueous phase was acidified with 1N HCl. This
was extracted twice with 20 mL EtOAc, and the combined organic
phases were washed with 50 mL 1N HCl. The organic phase was dried
over Na.sub.2SO.sub.4 filtered and evaporated to an oil, which
solidifies to yields 125 mg (38%) of the desired material. This
compound was used as such in the next step.
[0511] Step B. Preparation of (2S,2S) Phosphoric acid
mono-(2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-{2-[(morpholine-4-ca-
rbonyl)-amino]-3-naphthalen-1-yl-propionylamino}-hexyl) ester
(PL-498)
[0512] This compound was made as for the preparation of the product
of example 3 (step D) with 100 mg of
(2S)-2-[(morpholine-4-carbonyl)-amino]-3-naphthalen-1-yl-propionic
acid (step A of this example). The resulting precipitated residue
was further purified by reverse phase preparative HPLC. Yields 41
mg of the final compound.
[0513] LC-MS: 734.8 (M+H).sup.+, 95% pure.
[0514] .sup.1H NMR (CD.sub.3OD): .delta. 0.83-0.98 (m, 8H),
1.00-1.25 (m, 4H), 1.45-1.52 (m, 1H), 1.52-1.66 (m, 1H), 1.88-1.99
(m, 1H), 2.77-2.92 (m, 2H), 2.98-3.16 (m, 3H), 3.40-3.49 (m, 1H),
3.50-3.56 (m, 6H), 3.67-3.69 (m, 1H), 3.81-3.89 (m, 1H), 3.99-4.05
(m, 1H), 4.59 (t, J=6.0, 1H), 6.75 (d, J=8.0, 2H), 7.30-7.60 (m,
7H), 7.75 (d, J=8.0, 1H), 7.90 (d, J=7.8, 1H), 8.23 (d, J=7.8
2H).
[0515] .sup.31P NMR (CD.sub.3OD): .delta. 2.71
Example 5
Preparation of (2S,2S)-phosphoric acid
mono-{6-(2-acetylamino-3,3-diphenyl-propionylamino)-2-[(4-amino-benzenesu-
lfonyl)-isobutyl-amino]-hexyl}ester (PL-504)
[0516] Step A. Preparation
(2S)-2-acetylamino-3,3-diphenyl-propionic acid
[0517] To a solution of L-diphenylalanine (100 mg, 0.4 mmol)
(Peptech Corp.) in 5 mL 1N NaOH and 0.5 mL saturated
Na.sub.2CO.sub.3 (resulting solution at pH 10) was added acetyl
chloride (0.5 mmol) dissolved in 5 mL. Afterwards, the reaction
mixture was stirred at room temperature for 2 h. The alkaline
solution was extracted once with ether (10 mL) and the aqueous
phase was acidified with 1N HCl. This was extracted twice with 20
mL EtOAc, and the combined organic phases were washed with 50 mL 1N
HCl. The organic phase was dried over Na.sub.2SO.sub.4 filtered and
evaporated to an oil, which solidifies to yields 70 mg (60%) of the
desired material. This crude intermediate was used as such in the
next step.
[0518] Step B. Preparation of (2S,2S)-phosphoric acid
mono-{6-(2-acetylamino-3,3-diphenyl-propionylamino)-2-[(4-amino-benzenesu-
lfonyl)-isobutyl-amino]-hexyl}ester (PL-504)
[0519] This compound was made as for the preparation of the product
of example 3 (step D) with 100 mg of
(2S)-2-acetylamino-3,3-diphenyl-propionic acid (this example step
A). The final product was obtained in 30% yield (30 mg).
[0520] LC-MS: 689.3 (M+H).sup.+, 95% pure.
[0521] .sup.1H NMR (CD.sub.3OD): .delta. 0.77-1.04 (m, 9H),
1.10-1.17 (m, 1H), 1.23-1.49 (m, 1H), 1.46-1.57 (m, 1H), 1.78 (s,
3H), 1.88-1.99 (m, 1H), 2.80-2.92 (m, 2H), 2.92-3.08 (m, 2H),
3.63-3.75 (m, 1H), 3.79-3.95 (m, 1H), 4.00 (m, 1H), 4.34 (d,
J=11.3, 1H), 5.19-5.28 (m, 1H), 6.77-6.85 (m, 2H), 7.10-7.20 (m,
2H), 7.27-7.33 (m, 6H), 7.32-7.41 (m, 2H), 7.49-7.62 (m, 2H).
[0522] .sup.31P NMR (CD.sub.3OD): .delta. 2.70
Example 6
Preparation of
(1S,5S)-(1-{5-[(4-amino-3-fluoro-benzenesulfonyl)-isobutyl-amino]-6-phosp-
honooxy-hexylcarbamoyl)-2,2-diphenyl-ethyl)-carbamic Acid Methyl
Ester (PL-515)
[0523] First methodology: The preparation of the title compound is
based on scheme 3 of this invention.
[0524] Step A. Preparation of
(1-{5-[(4-amino-3-fluoro-benzenesulfonyl)-isobutyl-amino]-6-hydroxy-hexyl-
carbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl ester (X)
(PL-337)
[0525] The product of example 1, step F (0.624 g, 1 mmol) is
dissolved in 5 mL MeCN at 24.degree. C. SelectFluor 0.35 g (1 mmol)
is added in one portion and stirred for 1 h. 1 mL of water is added
and the solution was injected directly into a preparative
reverse-phase HPLC. The product was collected and lyophilized to
give 250 mg (38%) yield of a white solid.
[0526] LC-MS: 643.3 (M+H).sup.+, 99% pure.
[0527] .sup.1H NMR (MeOD): .delta. 0.71-0.85 (m 2H), 0.88 (d,
J=6.3, 6H), 0.91-0.96 (m, 2H), 1.21-1.29 (m, 1H), 1.41-1.52 (m, 1H)
1.82-1.92 (m, 1H), 2.61-2.68 (m, 1H), 2.81-2.85 (m, 2H), 2.94-3.05
(m, 2H), 3.38-3.40 (t, J=5, 1H), 3.49-3.52 (m, 5H), 4.28 (d, J=10,
1H), 4.87 (d, J=10, 1H) 6.90 (t, J=8.3, 1H), 7.20 (m, 2H), 7.28 (m,
3H), 7.33 (m, 3H), 7.39 (m, 4H).
[0528] Step B. Preparation of
(1S,5S)-(1-[5-[(4-amino-3-fluoro-benzenesulfonyl)-isobutyl-amino]-6-(diet-
hoxy-phosphoryloxy)-hexylcarbamoyl]-2,2-diphenyl-ethyl)carbamic
acid methyl ester
[0529] The product of step A was phosphorylated with
chlorodiethylphosphate following the procedure described in example
1, step G. Yields 157 mg, 68%.
[0530] LC-MS: 779.3 (M+H).sup.+, 95% pure.
[0531] .sup.1H NMR (CD.sub.3OD): .delta. 0.82 (m, 1H), 0.92 (d,
J=6.2, 8H), 0.96 (m, 3H), 1.36 (d, J=3.7, 6H), 1.90 (m, 1H), 2.69
(m, 1H), 2.89 (m, 1H), 2.98 (m, 2H), 3.56 (s, 3H), 3.74 (m, 1H),
3.93 (m, 1H), 4.03 (m, 1H), 4.12 (q, J=7.5 and 14.8, 4H), 4.32 (d,
J=11.4, 1H), 4.92 (d, J=11.4, 1H), 6.90 (t, J=8.3, 1H), 7.20 (m,
2H), 7.28 (m, 3H), 7.33 (m, 3H), 7.39 (m, 4H).
[0532] .sup.31P NMR (CD.sub.3OD): .delta. 1.65
[0533] Step C. Preparation of
(1S,5S)-(1-{5-[(4-amino-3-fluoro-benzenesulfonyl)-isobutyl-amino]-6-phosp-
honooxy-hexylcarbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl
ester (XI) (PL-515)
[0534] Deprotection was effected using the procedure described in
example 1, step G. Yields 101 mg.
[0535] LC-MS: 723.2 (M+H).sup.+, 95% pure.
[0536] .sup.1H NMR (CD.sub.3OD): .delta. 0.65-0.77 (m, 1H),
0.77-0.85 (m, 1H), 0.85-1.05 (m, 9H), 1.25-1.39 (m, 1H), 1.40-1.52
(m, 1H), 1.82-1.98 (m, 1H), 2.58-2.72 (m, 1H), 2.82-2.92 (m, 1H),
2.92-3.05 (m, 2H), 3.54 (s, 3H), 3.64-3.75 (m, 1H), 3.80-3.92 (m,
1H), 3.91-4.04 (m, 1H), 4.29 (d, J=11.4, 1H), 7.19 (t, J=6.6, 1H),
7.13-7.21 (m, 2H), 7.22-7.33 (m, 6H), 7.34-7.38 (m, 2H), 7.39-7.48
(m, 2H).
[0537] .sup.31P NMR (CD.sub.3OD): .delta. 2.74
[0538] Second methodology: The preparation of the title compound is
based on scheme 4 of this invention.
[0539] Step A. Preparation
(1S,5S)-(1-(5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-phosphonooxy-h-
exylcarbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl ester
(PL-461)
[0540] (2S)-2-methoxycarbonylamino-3,3-diphenyl-propionic acid
((example 1, step E) 0.9 g, 3 mmol) was activated in DMF (5 mL)
with EDAC (1.7 g, 9 mmol) and HOBt (1.2 g, 9 mmol). To the solution
was added 1.17 g of (2S)-phosphoric acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
diethyl ester (IX) (example 3, step B) and the mixture stirred for
3 h. 20 g of Amberlite XAD-2 resin was then added and the beads
were left to soak for 10 min. The resin was transferred into a
glass filter and washed thoroughly with distilled water (400 mL)
and 200 mL of 1M NaHCO.sub.3. The beads were then washed with
4.times.50 ml portions of MeOH then EtOAc 200 mL. The organic phase
was evaporated. The residue was adsorbed onto silica gel and passed
through a short silica gel column (EtOAc) to yield 2.4 g (83%) of
white solid after evaporation.
[0541] NMR identical as in example 1, step H.
[0542] Step B. Preparation
(1S,5S)-{1-[5-[(4-amino-3-fluoro-benzenesulfonyl)-isobutyl-amino]-6-(diet-
hoxy-phosphoryloxy)-hexylcarbamoyl]-2,2-diphenyl-ethyl}-carbamic
acid methyl ester (XII)
[0543] The product of step A above,
(1S,5S)-(1-{5-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-phosphonooxy
hexylcarbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl ester
(0.555 g, 0.73 mmol) was dissolved in 5 mL MeCN. Selectfluor (0.26
g, 0.7 mmol) was added and the mixture stirred for 30 min. The
mixture was purified by reverse phase preparative HPLC and
lyophilized to yield 278 mg (48% yield) white solid.
[0544] .sup.1H NMR identical as previous entry, see first
methodology above.
[0545] Step C. Preparation
(1S,5S)-(1-{5-[(4-amino-3-fluoro-benzenesulfonyl)-isobutyl-amino]-6-phosp-
honooxy-hexylcarbamoyl}-2,2-diphenyl-ethyl)-carbamic acid methyl
ester (XIII, in this specific case is compound XI) (PL-515)
[0546] The procedure make this derivative was as described in the
deprotection step for the methodology above. Yields 139 mg 70%
after reverse phase HPLC.
[0547] .sup.1H NMR identical as previous entry, see first
methodology above.
Example 7
Preparation of (2S,2S)-acetic acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl Ester (PL-521)
[0548] The preparation of the title derivative is based on scheme 5
of this invention.
[0549] Step A. Preparation of (2S)-acetic acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesulfonyl-
)-isobutyl-amino]-hexyl ester (XIV, R.sub.1A.dbd.CH.sub.3)
[0550] To a stirred solution of
(1S)-{4-[(5-tert-butoxycarbonylamino-1-hydroxymethyl-pentyl)-isobutyl-sul-
famoyl]-phenyl}carbamic acid tert-butyl ester (intermediate product
(VII) of example 1, step D, 97 mg, 0.18 mmol) in anhydrous
CH.sub.2Cl.sub.2 (3 mL) was added N,N-dimethylaminopyridine (22 mg,
0.18 mmol) and acetic anhydride (0.014 mL, 0.18 mmol). The mixture
was stirred at room temperature for 1 hour. The solvent was
evaporated. Ethyl acetate (50 mL) was added and the organic layer
was washed with water (30 mL), then dried with Na.sub.2SO.sub.4 and
concentrated. The residue was purified by flash chromatography
eluting with ethyl acetate. The yield obtained was quantitative
(100 mg).
[0551] LC-MS: 586.2 (M+H).sup.+, 95% pure
[0552] Step B. Preparation of (2S)-acetic acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
(XV, R.sub.1A.dbd.CH.sub.3)
[0553] This derivative was prepared from (2S)-acetic acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesulfonyl-
)-isobutyl-amino]-hexyl ester as described in example 15, step B.
The yellow solid (66 mg) was used for the next reaction without
purification.
[0554] LC-MS: 386.2 (M+H).sup.+, 95% pure
[0555] Step C. Preparation of (2S,2S)-acetic acid
2-[(4-amino-benzenesulfonyl-)isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl ester (XVI, R.sub.1A.dbd.CH.sub.3)
(PL-521)
[0556] This derivative was prepared from (2S)-acetic acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
(product of step B) as described in example 15, step B. The final
product was purified by flash chromatography with a mixture of
eluents hexane/ethyl acetate (2/8). A yellow solid was obtained in
70% yield (70 mg).
[0557] LC-MS: 667.3 (M+H).sup.+, 95% pure
[0558] .sup.1H NMR(acetone-d.sub.6): .delta. 0.85-0.97 (m, 12H),
1.21-1.41 (m, 2H), 1.88-2.00 (s, 3H), 2.59-2.69 (m, 1H), 2.83-2.90
(m, 1H), 2.90-3.01 (m, 1H), 3.01-3.10 (br s, 1H), 3.45-3.60 (s,
3H), 3.70-3.80 (m, 1H), 3.93-4.00 (m, 1H), 4.00-4.11 (m, 1H),
4.38-4.45 (d, J=11.0, 1H), 4.89-4.98 (t, J=10.0, 1H), 5.43-5.58 (br
s, 1H), 6.28-6.48 (d, J=8.9, 11-1), 6.72-6.83 (d, J=8.0, 2H),
6.85-6.93 (br s, 1H), 7.12-7.22 (t, J=7.4, 1H), 7.21-7.31 (d,
J=7.0, 4H), 7.31-7.45 (m, 5H), 7.48-7.57 (d, J=8.0, 2H).
Example 8
Preparation of (2S,2S)-nicotinic acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl Ester (PL-520)
[0559] Step A. Preparation of (2S)-nicotinic acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesulfonyl-
)-isobutyl-amino]-hexyl ester (XIV, R.sub.1A=3-pyridyl)
[0560]
(1S)-{4-[(5-tert-butoxycarbonylamino-1-hydroxymethyl-pentyl)-isobu-
tyl-sulfamoyl]-phenyl}-carbamic acid tert-butyl ester (intermediate
product (VII) of example 1, step D, 130 mg, 0.24 mmol) was
dissolved in anhydrous DMF (1 mL) and treated with 0.066 mL (0.48
mmol) of triethylamine followed by EDC (120 mg, 0.65 mmol), HOBt
(88 mg, 0.65 mmol) and nicotinic acid (27 mg, 0.22 mmol). The
mixture was stirred overnight at room temperature. The product was
extracted with ethyl acetate (40 mL) and water (40 mL). The organic
phase was separated and dried with Na.sub.2SO.sub.4, then
evaporated to give 200 mg of crude product. This compound was
purified by flash chromatography with ethyl acetate as the eluent.
A clear oil was obtained in 100% yield (150 mg).
[0561] LC-MS: 649.3 (M+H).sup.+, 95% pure
[0562] .sup.1H NMR (acetone-d.sub.6): .delta. 0.90-1.14 (d, J=5.9,
6H), 1.31-1.42 (m, 2H), 1.48 (s, 9H), 1.51-1.55 (m, 2H), 1.59 (s,
9H), 1.62-1.69 (m, 1H), 1.72-1.83 (m, 1H), 3.00-3.11 (m, 2H),
3.11-3.17 (m, 1H), 3.19-3.27 (m, 1H), 4.15-4.24 (m, 1H), 4.35-4.44
(t, J=9.1, 1H), 4.50-4.58 (dd, J=4.4 and 11.5, 1H), 5.89-5.99 (br
s, 1H), 7.53-7.60 (m, 1H), 7.70-7.77 (d, J=8.2, 2H), 7.80-7.87 (d,
J=8.2, 2H), 8.24-8.31 (d, J=7.3, 1H), 8.75-8.82 (m, 1H), 8.82-8.88
(m, 1H), 9.12-9.18 (br s, 1H).
[0563] Step B. Preparation of (2S)-nicotinic acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
(XV, R.sub.1A=3-pyridyl)
[0564] The product of step A, (2S)-nicotinic acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesulfonyl-
)-isobutyl-amino]-hexyl ester (150 mg, 0.23 mmol), was dissolved in
CH.sub.2Cl.sub.2 (5 mL) and trifluoroacetic acid (1 mL) was added.
The mixture was stirred during 2 hours at room temperature. The
solvent was evaporated and the residue was extracted with ethyl
acetate (40 mL) and NaOH 1M (40 mL) (pH=10). The organic portion
was separated, dried with Na.sub.2SO.sub.4 and evaporated. The
residue (100 mg) was used for the next reaction without further
purification. The yield was quantitative.
[0565] LC-MS: 449.2 (M+H).sup.+, 95% pure
[0566] Step C. Preparation of (2S,2S)-nicotinic acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl ester (PL-520)
[0567] The product of step B, (2S)-nicotinic acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
(100 mg, 0.22 mmol) was dissolved in anhydrous DMF (2 mL) and
treated with 0.062 mL (0.45 mmol) of triethylamine followed by EDC
(100 mg, 0.56 mmol), HOBt (75 mg, 0.56 mmol) and
(2S)-2-methoxycarbonylamino-3,3-diphenyl-propionic acid (56 mg,
0.19 mmol). The mixture was stirred overnight at room temperature.
The product was extracted with ethyl acetate (40 mL) and water (40
mL). The organic layer was separated and dried with
Na.sub.2SO.sub.4, then evaporated to give 160 mg of crude oil. The
residue was purified by flash chromatography with a mixture of
eluents hexane/ethyl acetate (2/8). The title compound was obtained
as a clear oil in 20% yield (25 mg).
[0568] LC-MS: 730.2 (M+H).sup.+, 95% pure
[0569] .sup.1H NMR (acetone-d.sub.6): .delta. 0.80-0.97 (m, 9H),
0.97-1.13 (m, 2H), 1.26-1.40 (m, 1H), 1.40-1.57 (m, 1H), 2.61-2.73
(m, 1H), 2.86-2.98 (m, 2H), 3.00-3.17 (m, 2H), 3.45-3.59 (s, 3H),
3.91-4.00 (m, 1H), 4.24-4.34 (m, 1H), 4.34-4.47 (m, 2H), 4.90-4.99
(t, J=9.7, 1H), 6.35-6.44 (m, 1H), 6.68-6.79 (d, J=7.9, 1H),
6.91-7.00 (br s, 1H), 7.13-7.22 (m, 2H), 7.22-7.31 (m, 3H),
7.35-7.48 (m, 4H), 7.49-7.64 (m, 2H), 7.75-7.84 (m, 1H), 8.25-8.36
(m, 1H), 8.76-8.88 (br s, 1H), 9.12-9.26 (br s, 1H).
Example 9
Preparation of (2S,2S)-dimethylamino-acetic acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl Ester (PL-534)
[0570] Step A. Preparation of (2S)-dimethylamino-acetic acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesulfonyl-
)-isobutyl-amino]-hexyl ester (XIV,
R.sub.1A=(CH.sub.3).sub.2NCH.sub.2--)
[0571] This title compound was obtained from
(1S)-{4-[(5-tert-butoxycarbonylamino-1-hydroxymethyl-pentyl)-isobutyl-sul-
famoyl]-phenyl}-carbamic acid tert-butyl ester (intermediate
product (VII) of example 1, step D) as described example 15, step A
using N,N-dimethylglycine. The clear oil was obtained in 100% yield
(150 mg).
[0572] LC-MS: 629.3 (M+H).sup.+, 95% pure
[0573] .sup.1H NMR (acetone-d.sub.6): .delta. 0.81-0.95 (d, J=6.1,
6H), 1.18-1.30 (m, 2H), 1.32-1.43 (s, 9H), 1.43-1.52 (s, 8H),
1.52-1.62 (m, 1H), 1.93-2.00 (m, 1H), 2.19-2.29 (s, 4H), 2.69-2.80
(m, 4H), 2.90-3.05 (m, 6H), 3.60-3.65 (m, 1H), 3.85-3.97 (m, 1H),
3.98-4.08 (m, 1H), 4.08-4.14 (m, 1H), 5.78-5.88 (m, 1H), 7.68-7.80
(m, 3H), 8.80-8.88 (br s, 1H).
[0574] Step B. Preparation of (2S)-dimethylamino-acetic acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
(XV, R.sub.1A.dbd.(CH.sub.3).sub.2NCH.sub.2--)
[0575] The title derivative was prepared from
(2S)-dimethylamino-acetic acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesul-
fonyl)-isobutyl-amino]-hexyl ester as described in example 15, step
B. The final product (100 mg) was used as such in the next
step.
[0576] LC-MS: 429.3 (M+H).sup.+, 90% pure
[0577] Step C. Preparation of (2S,2S)-dimethylamino-acetic acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl ester (PL-534)
[0578] This title compound was prepared from
(2S)-dimethylamino-acetic acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester as
described in example 15, step C. The crude product was purified by
LC-preparative. The final compound was obtained in 10% yield (10
mg).
[0579] LC-MS: 710.3 (M+H).sup.+, 92% pure
[0580] .sup.1H NMR (acetone-d.sub.6): .delta. 0.81-0.98 (m, 12H),
1.14-1.30 (m, 2H), 1.31-1.45 (m, 1H), 2.58-2.77 (m, 2H), 2.79-2.90
(m, 2H), 3.42-3.56 (s, 3H), 3.75-3.85 (m, 1H), 3.99-4.17 (m, 3H),
4.23-4.35 (m, 1H), 4.36-4.45 (m, 1H), 4.86-4.96 (m, 1H), 6.33-6.42
(m, 1H), 6.74-6.83 (m, 1H), 6.85-6.90 (m, 1H), 7.12-7.22 (m, 3H),
7.23-7.31 (m, 4H), 7.31-7.44 (m, 5H), 7.47-7.55 (m, 1H), 7.73-7.80
(m, 1H).
Example 10
Preparation of (2S,2S)-2-amino-3-methyl-butyric acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl Ester (PL-530)
[0581] Step A. Preparation of
(2S)-2-benzyloxycarbonylamino-3-methyl-butyric acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesulfonyl-
)-isobutyl-amino]-hexyl ester (XIV,
R.sub.1A.dbd.(CH.sub.3).sub.2CHCH(NH.sub.2)--)
[0582] This title compound was obtained from
(1S)-{4-[(5-tert-butoxycarbonylamino-1-hydroxymethyl-pentyl)-isobutyl-sul-
famoyl]-phenyl}-carbamic acid tert-butyl ester (intermediate
product (VII) of example 1, step D) as described in example 15,
step A using (2S)-2-benzyloxycarbonylamino-3-methyl-butyric acid.
The crude product was purified by flash chromatography eluting with
a mixture of hexane/ethyl acetate (1/1). The yield obtained was
100% (150 mg).
[0583] LC-MS: 777.3 (M+H).sup.+, 95% pure
[0584] .sup.1H NMR (acetone-d.sub.6): .delta. 0.80-1.00 (m, 14),
1.13-1.28 (s, 2H), 1.30-1.44 (s, 11H), 1.45-1.56 (s, 10), 1.58-1.67
(m, 1H), 2.87-3.04 (m, 4H), 3.84-3.97 (m, 1H), 3.97-4.12 (m, 2H),
4.12-4.21 (m, 1H), 4.99-5.14 (m, 2H), 5.78-5.89 (m, 1H), 6.38-6.52
(m, 1H), 7.24-7.34 (m, 1H), 7.34-7.41 (m, 2H), 7.65-7.83 (m, 4H),
8.77-8.86 (m, 1H).
[0585] Step B. Preparation of
(2S)-benzyloxycarbonylamino-3-methyl-butyric acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
(XV, R.sub.1A.dbd.(CH.sub.3).sub.2CHCH(NH.sub.2)--)
[0586] This derivative was prepared from
(2S)-2-benzyloxycarbonylamino-3-methyl-butyric acid
6-tert-butoxycarbonylamino-2-[(4-tert-butoxycarbonylamino-benzenesulfonyl-
)-isobutyl-amino]-hexyl ester (product of step A) as described in
example 15, step B. The final compound was obtained in quantitative
yield (110 mg) and used for the next step without purification.
[0587] LC-MS: 577.3 (M+H).sup.+, 90% pure
[0588] Step C. Preparation of
(2S,2S)-2-benzyloxycarbonylamino-3-methyl-butyric acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl ester
[0589] The title compound was obtained from
(2S)-benzyloxycarbonylamino-3-methyl-butyric acid
6-amino-2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-hexyl ester
(product of step B) as described in example 15, step C. The clear
oil was obtained in 86% yield (120 mg).
[0590] LC-MS: 858.3 (M+H).sup.+, 95% pure
[0591] Step D. Preparation of (2S,2S)-2-amino-3-methyl-butyric acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl ester (PL-530)
[0592] To a stirred solution of
(2S,2S)-2-benzyloxycarbonylamino-3-methyl-butyric acid
2-[(4-amino-benzenesulfonyl)-isobutyl-amino]-6-(2-methoxycarbonylamino-3,-
3-diphenyl-propionylamino)-hexyl ester (step C, 120 mg, 0.14 mmol)
in anhydrous THF (8 mL), under nitrogen atmosphere, was added
palladium 10% wt. on activated carbon (160 mg). The mixture was
reacted under hydrogen atmosphere overnight, at room temperature.
The solution was filtered and the palladium on carbon was washed
with THF (50 mL). The solvent was evaporated and the residue (110
mg) was purified by flash chromatography using ethyl acetate as the
eluent. The clear oil was obtained in 47% yield (47 mg).
[0593] LC-MS: 796.4 (M+H).sup.+, 95% pure
[0594] .sup.1H NMR (acetone-d.sub.6): .delta. 0.84-0.97 (m, 12H),
0.97-1.08 (m, 2H), 1.27-1.43 (m, 3H), 1.49-1.62 (m, 4H), 1.80-1.93
(m, 1H), 1.94-2.00 (m, 1H), 2.36-2.46 (m, 1H), 2.58-2.74 (m, 2H),
2.86-2.96 (m, 3H), 2.99-3.10 (m, 2H), 3.46-3.52 (s, 3H), 3.52-3.60
(m, 2H), 3.75-3.87 (m, 2H), 3.95-4.04 (m, 1H), 4.10-4.18 (m, 1H),
4.37-4.44 (m, 1H), 4.89-4.97 (m, 1H), 5.40-5.48 (m, 1H), 6.30-6.40
(m, 1H), 6.76-6.83 (d, J=8.2, 1H), 6.87-7.03 (m, 2H), 7.14-7.22 (m,
1H), 7.23-7.34 (m, 3H), 7.35-7.45 (m, 4H), 7.50-7.56 (m, 1H),
7.57-7.65 (m, 1H).
Example 11
Inhibitory Activity and Cytotoxicity
[0595] The active ingredients which are released from the
Lysine-based compounds described herein have been shown to inhibit
HIV aspartyl protease (U.S. Pat. No. 6,632,816). After oral dosing,
plasma levels of the Lysine-based compound; PL-461 in rats are
insignificant or under detection limits, compared to plasma levels
of the active ingredient; PL-100, indicating that the phosphate
moiety of the Lysine-based compound (e.g., PL-461) is metabolically
cleaved and majority of this compound is converted to the
corresponding active ingredient (e.g., PL-100) in vivo.
[0596] The cross-resistance profile of a representative active
ingredient; PL-100 against 63 HIV isolates with reduced
susceptibility to all approved protease inhibitors (PIs) including
atazanavir is tested herein.
[0597] Briefly, the antiviral activity of PL-100 was determined
using MT4 cells infected with a laboratory adapted HIV strain
(NL4-3). For comparison, seven approved protease inhibitors (PIs)
were tested in parallel. Result of this experiment is presented in
Table 3.
[0598] These data suggest that the antiviral activity of PL-100 is
comparable to that of the other PIs.
[0599] Cytotoxicity of PL-100 and other P is was evaluated in the
same cell culture system. The selectivity index (SI) is the ratio
of cytotoxicity (CC.sub.50) to antiviral activity (EC.sub.50). The
selective index is similar to the therapeutic index but relates to
in vitro studies.
[0600] This result indicates that PL-100 is a potent, selective and
non-cytotoxic PI. TABLE-US-00001 TABLE 3 Protease SI Inhibitor
EC.sub.50 (nM) CC.sub.50 (nM) (CC.sub.50/EC.sub.50) Atazanavir 4
55,000 13,750 Amprenavir 47 >100,000 >2,128 Indinavir 67
>100,000 >1,493 Lopinavir 19 28,000 1,474 Nelfinavir 29 8,000
276 Ritonavir 61 25,000 410 Saquinavir 12 19,000 1,583 PL-100 16
33,000 2,063
[0601] Atazanavir (ATV) is manufactured by BMS (Bristol Myers
Squibb), Amprenavir (APV) is manufactured by GSK (Glaxo
SmithKline), Indinavir (IDV) is manufactured by Merck, Lopinavir
(LPV) is manufactured by Abott, Nelfinavir (NFV) is manufactured by
Pfizer, Ritonavir (RTV) is manufactured by Abott, Saquinavir (SQV)
is manufactured by Roche.
[0602] HIV PI class inhibitors may sometimes demonstrate binding to
plasma protein which may affect their potency in vivo. The effect
of 40% human serum on the antiviral activity of PL-100 in MT4 cells
infected with NL4-3 was therefore evaluated. As shown in Table 4,
the addition of human serum reduced the antiviral activity of
PL-100 by 6-fold, which result in an increase of its EC.sub.50 to
100 nM. This is similar to what is reported for the approved drugs
nelfinavir (NFV) and indinavir (IDV). So protein binding of PL-100
is not expected to be a significant issue in the clinic.
TABLE-US-00002 TABLE 4 EC.sub.50 nM (Fold Culture condition Change)
10% FBS* 18 (1.0) 10% FBS* + 40% human 106 (5.9) serum *Fetal
bovine serum
Example 12
Cross-Resistance Profile
[0603] The cross-resistance profile of PL-100 against 63 HIV
isolates with reduced susceptibility to all approved protease
inhibitors (P is) including atazanavir was thus tested.
[0604] The panel of 63 viral strains was selected based on the
following rationale: 1) High-level loss of susceptibility to
specific P is; 2) High-level loss of susceptibility to multiple P
is; 3) Good representation of the primary mutations. This panel
consists of resistant viruses from highly PI-experienced patients
with high-level PI resistance.
[0605] The genotype of these viruses encompasses a wide variety of
mutational patterns. We particularly looked for the following
primary PI mutations: D30N, L33F/I, M461/L, G48V, I50L/V,
V82A/F/S/T, I84V and L90M as defined by the International AIDS
Society (IAS)-USA. Table 5 illustrates the repartition and the
strength of the chosen panel. TABLE-US-00003 TABLE 5 No. of primary
PI No. of viral mutations strains 0 3 1 6 2 11 3 22 4 17 5 3 6
1
[0606] The mutation mentioned above refers to original (wild type)
amino acid (one letter code)/position in the HIV-1
protease/resulting amino acid change(s) (one letter code). The one
letter code amino acid is known in the art.
[0607] For example: D30N relates to: aspartic acid (D) found at
position 30 of HIV-1 protease which has been changed (mutated) for
an asparagine (N):
[0608] L33F/I relates to: leucine (L) found at position 33 of HIV-1
protease which has been changed (mutated) for either phenylalanine
(F) or isoleucine (I); etc.,
[0609] Table 6, illustrates the result of the cross-resistance
profile of 63 diverse resistant strains. TABLE-US-00004 TABLE 6
Fold change in IC50 vs. Reference SAMPLE ID Key Mutations.sup.1
Panel ID ATV AMP IDV LPV NFV SQV PL-100 O3-129572
24I/33F/54V/82A/84V 1 ##STR101## ##STR102## ##STR103## ##STR104##
##STR105## ##STR106## O3-129573 24I/33F/46I/54L/82A 3 ##STR107##
##STR108## ##STR109## ##STR110## 1.5 ##STR111## O3-129574
54V/82A/84V/90M 5 ##STR112## ##STR113## ##STR114## ##STR115##
##STR116## ##STR117## O3-129575 33F/54V/73S/82A/84V/90M 6
##STR118## ##STR119## ##STR120## ##STR121## ##STR122## ##STR123##
O3-129576 33F/46L/54V/82A/84V/90M 7 ##STR124## ##STR125##
##STR126## ##STR127## ##STR128## ##STR129## O3-129578 46I/82T/84V
11 ##STR130## ##STR131## ##STR132## 1.6 1 1.2 O3-129579 46I/82T/84V
13 ##STR133## ##STR134## ##STR135## ##STR136## ##STR137##
##STR138## O3-129580 46I/82T/84V/90M 15 ##STR139## ##STR140##
##STR141## ##STR142## ##STR143## ##STR144## O3-129581
33I/46I/84V/88D/90M 16 ##STR145## ##STR146## ##STR147## ##STR148##
##STR149## ##STR150## O3-1129583 84V/90M 21 ##STR151## ##STR152##
##STR153## ##STR154## ##STR155## 2.3 O3-129584 33F/50V 24
##STR156## 1.3 ##STR157## ##STR158## 1.7 ##STR159## O3-129585
48V/54V/82A/90M 26 ##STR160## ##STR161## ##STR162## ##STR163##
##STR164## 1.9 O3-129586 30N/88D/90M 28 1.8 2.3 1.2 ##STR165##
##STR166## 1.4 O3-148709.sup.2 32I/47A 1 1 ##STR167## ##STR168##
##STR169## ##STR170## 0.2 1.4 O4-148710.sup.2 46I/47V/84V 118
##STR171## ##STR172## ##STR173## ##STR174## ##STR175## ##STR176##
##STR177## O4-148711 46I/47A/84V 119 ##STR178## ##STR179##
##STR180## ##STR181## ##STR182## 1.1 ##STR183## O4-148712 46I/88S
122 ##STR184## 0.3 ##STR185## 1.7 ##STR186## 1.9 0.8 O4-148713
46I/88S 123 ##STR187## 0.2 2 0.7 4.6 0.6 0.4 O4-148712 46I/88S 124
##STR188## 0.9 ##STR189## ##STR190## ##STR191## 2 1.1 O4-148715
33F/54L/88S/90M 9 ##STR192## ##STR193## ##STR194## ##STR195##
##STR196## ##STR197## ##STR198## O4-148716 33F/54L/88S/90M 10
##STR199## ##STR200## ##STR201## ##STR202## ##STR203## ##STR204##
1.7 O4-148718 33I/46I/84V/88D/90M 19 ##STR205## ##STR206##
##STR207## ##STR208## ##STR209## ##STR210## ##STR211## O4-148719
33F/46I/84V/88D/90M 20 ##STR212## ##STR213## ##STR214## ##STR215##
##STR216## ##STR217## ##STR218## O4-148720 30N/33F/46L/54L/84V/88D
125 ##STR219## ##STR220## ##STR221## ##STR222## ##STR223##
##STR224## ##STR225## O4-148721 32I/46I/47V/50L 127 ##STR226##
##STR227## 2.1 ##STR228## ##STR229## 0.5 1.1 O4-148722
33F/54L/82A/90M 40 ##STR230## ##STR231## ##STR232## ##STR233##
##STR234## ##STR235## ##STR236## O4-148723 33F/54L/82A/90M 42
##STR237## ##STR238## ##STR239## ##STR240## ##STR241## ##STR242##
##STR243## O4-148724 33F/54V/73S/82A/90M 44 ##STR244## ##STR245##
##STR246## ##STR247## ##STR248## ##STR249## ##STR250## O4-148725
32I/46L/47V/84V 53 ##STR251## ##STR252## ##STR253## ##STR254##
##STR255## 0.8 0.6 O4-148726 33F/54L/82A/84V 132 ##STR256##
##STR257## ##STR258## ##STR259## ##STR260## ##STR261## ##STR262##
O4-148727 33F/54V/82A/84V 133 ##STR263## ##STR264## ##STR265##
##STR266## ##STR267## ##STR268## ##STR269## O4-148728
33F/46I/53L/82A/84V 134 ##STR270## ##STR271## ##STR272## ##STR273##
##STR274## ##STR275## ##STR276## O4-148729 33F/54V/82T/84V/90M 61
##STR277## ##STR278## ##STR279## ##STR280## ##STR281## ##STR282##
##STR283## O4-148730 33F/46L/53L/54V/82A 67 ##STR284## ##STR285##
##STR286## ##STR287## ##STR288## ##STR289## ##STR290## O4-148731
46L/54M/82L/84V/90M 69 ##STR291## ##STR292## ##STR293## ##STR294##
##STR295## ##STR296## ##STR297## O4-148732 46L/54V/82A/90M 74
##STR298## ##STR299## ##STR300## ##STR301## ##STR302## ##STR303##
##STR304## O4-148733 46I/54V/82A/84V 76 ##STR305## ##STR306##
##STR307## ##STR308## ##STR309## ##STR310## ##STR311## O4-148734
46L/54V/73C/84V/90M 78 ##STR312## ##STR313## ##STR314## ##STR315##
##STR316## ##STR317## ##STR318## O4-148735 46I/82T/84V/90M 81
##STR319## ##STR320## ##STR321## ##STR322## ##STR323## ##STR324##
##STR325## O4-148736 46I/82A/84V/90M 82 ##STR326## ##STR327##
##STR328## ##STR329## ##STR330## ##STR331## ##STR332## O4-148737
54V/82A/84V/90M 84 ##STR333## ##STR334## ##STR335## ##STR336##
##STR337## ##STR338## ##STR339## O4-148738 33F/54V/82A/84V/90M 86
##STR340## ##STR341## ##STR342## ##STR343## ##STR344## ##STR345##
##STR346## O4-148739 48V/54V/82A/90M 138 ##STR347## 1.1 ##STR348##
##STR349## ##STR350## ##STR351## 1.1 O4-148740 46L/48V/82A/84V/90M
141 ##STR352## ##STR353## ##STR354## ##STR355## ##STR356##
##STR357## ##STR358## O4-148741 48V/82A/84V/90M 142 ##STR359##
##STR360## ##STR361## ##STR362## ##STR363## ##STR364## ##STR365##
O4-148742 46L/48V/82A/84V 94 ##STR366## ##STR367## ##STR368##
##STR369## ##STR370## ##STR371## 1.2 O4-148743 46L/48V/82A/90M 95
##STR372## ##STR373## ##STR374## ##STR375## ##STR376## ##STR377##
##STR378## O4-148744 46L/50L/54V/82A 98 ##STR379## ##STR380##
##STR381## ##STR382## ##STR383## ##STR384## ##STR385## O4-148745
46I/50L/54V/82A 99 ##STR386## ##STR387## ##STR388## ##STR389##
##STR390## 2.2 ##STR391## O4-148746 33I/46I/84V/90M 105 ##STR392##
##STR393## ##STR394## ##STR395## ##STR396## ##STR397## ##STR398##
O4-148747 33I/46I/84V/90M 107 ##STR399## ##STR400## ##STR401##
##STR402## ##STR403## ##STR404## ##STR405## O4-148748 37S/41K/70E
115 1.4 0.7 1.2 1 1.6 1.2 0.9 O4-148749 37T/41K/70E 151 1.5 1.9 1.4
1.3 1.9 1.2 1.9 O4-148750 46I/53L/82T/90M 172 ##STR406## ##STR407##
##STR408## ##STR409## ##STR410## ##STR411## ##STR412## O4-148751
54V/84V 171 ##STR413## ##STR414## ##STR415## ##STR416## ##STR417##
##STR418## 0.8 O4-148752 54V/84V 152 ##STR419## ##STR420##
##STR421## ##STR422## ##STR423## ##STR424## 1.2
[0610] In Table 6 above, (.sup.1) indicates that mutations used in
selecting viruses in at least one group; mixtures were ignored for
this purpose, (.sup.2) indicates non-B subtype and (*) indicates
50% inhibition not reached at highest drug concentration tested and
that an arbitrary FC value (400) was therefore assigned.
[0611] The key mutations listed in Table 6, only refers to the
position in the HIV-1 protease which has been mutated and the
resulting amino acid (one letter code) found in the mutated
protease of the resistant strain. The original amino acid is not
indicated.
[0612] For example, 33F/46L/54V/82A/84V/90M (panel ID 7 of Table 6)
indicates that amino acid at position 33 has been mutated to
phenylalanine (F), amino acid at position 46 has been mutated to
leucine (L), amino acid at position 54 has been mutated to valine
(V), amino acid at position 82 has been mutated to alanine (A),
amino acid at position 84 has been mutated to valine (V) and amino
acid at position 90 has been mutated to methionine (M). Similarly
with the other resistant viral strains listed in Table 6.
[0613] The results presented in the above Table 6 indicates the
fold-change (FC) in IC.sub.50 vs the reference strain (NL-4.3). The
dark cells indicate a FC.ltoreq.10, the grey cells indicate FC
values of 2.5.ltoreq.FC<10 and the white cells indicate a FC
value of; 0.ltoreq.FC<2.5. The results of cross-resistance
profile are further illustrated in the graph of FIG. 1.
[0614] Table 7 summarizes the phenotypic susceptibility results
obtained against the 63 diverse, multi-PI-resistant strains
mentioned above. TABLE-US-00005 TABLE 7 ATV APV IDV LPV NFV SQV
PL-100 Median FC 15.6 9.7 8.1 17.9 15 23.2 3.6 Mean FC 25.7 18.5
16.5 31.3 31.7 85 8.7 % FC < 2.5 16 14 13 10 10 27 37 % FC <
10 38 52 54 37 27 40 76 % FC > 50 22 8 5 19 10 37 3
[0615] In the cross-resistance profiling studies against the 63
resistant strains mentioned herein, EC.sub.95 of PL-100 was
determined against each strain. The median EC.sub.95 represents the
concentration required to inhibit 95% of the viral replication of
50% diverse drug-resistant HIV-1 variants tested. Table 8
illustrates the percentage of resistant strains tested with
protein-binding adjusted EC.sub.95s.ltoreq.200 ng/ml. The resistant
strains are grouped by the number of primary mutations they have.
TABLE-US-00006 TABLE 8 No. of primary PI % viral strains with
mutations EC.sub.95s .ltoreq. 200 ng/ml 0 100 1 100 2 36 3 23 4 0
5-6 0
[0616] The EC.sub.95 of PL-100 was determined against each of the
63 resistant strains indicated herein. Table 9 illustrates the
percentage of resistant strains tested with protein-binding
adjusted EC.sub.95s.ltoreq.630 ng/ml. The resistant strains are
grouped by the number of primary mutations they have.
TABLE-US-00007 TABLE 9 No. of primary PI % viral strains with
mutations EC.sub.95s .ltoreq. 630 ng/ml 0 100 1 100 2 82 3 55 4 29
5-6 0
[0617] Results presented herein indicate that PL-100 has a
favourable cross-resistance profile against several HIV
isolates.
Example 13
Bioavailability of Compounds
[0618] In order to improve the chemical stability, aqueous
solubility of the active ingredients, various Lysine-based
compounds, such as PL-100 derivatives (PL-461, PL-462, etc.) were
designed, synthesized and purified (FIG. 2, Table 10). The active
ingredient has been shown to be efficient against an HIV-1 aspartyl
protease (U.S. Pat. No. 6,632,816). Also as mentioned herein, the
active ingredients present potent antiviral activity when tested on
non-mutated HIV-1 viral strain (NL4.3 as the wild type virus) as
well as several mutant strains.
[0619] Various PL-100 precursors were thus developed and tested.
Examples of representative PL-100 precursors are illustrated in
Table 10. PL-100 and PL-337 have been chosen herein as
representative active ingredient among those disclosed in U.S. Pat.
No. 6,632,816 to Stranix et al. for improving pharmacokinetics of
these potent inhibitors.
[0620] The compounds listed in Table 10 were prepared by following
scheme 1, 1A, 2, 3, 4 or 5; and more particularly as described in
each example listed above. The numbers of the compounds listed in
Table 10 (Ex. No.) corresponds to the example numbers presented
above. TABLE-US-00008 TABLE 10 Structures of exemplary lysine based
compounds and active ingredients encompassed by the present
invention I ##STR425## D, L, DL Ex. No R, S, (PL-#) X Y n R.sub.1
R.sub.2 R.sub.3 R.sub.6 X'/Y' RS 1 (PL-461) 4-NH.sub.2 H 4
(HO).sub.2P(O) (C.sub.6H.sub.5).sub.2CH CH.sub.3O--CO iso-butyl H/H
S, S 2 (PL-462) 4-NH.sub.2 H 4 (NaO).sub.2P(O)
(C.sub.6H.sub.5).sub.2CH CH.sub.3O--CO iso-butyl H/H S, S 3
(PL-507) 4-NH.sub.2 H 4 (HO).sub.2P(O) Naphthyl-2-CH.sub.2
CH.sub.3O--CO iso-butyl H/H S, S 4 (PL-498) 4-NH.sub.2 H 4
(HO).sub.2P(O) Naphthyl-1-CH.sub.2 4-morpholine- iso-butyl H/H S, S
CO 5 (PL-504) 4-NH.sub.2 H 4 (HO).sub.2P(O)
(C.sub.6H.sub.5).sub.2CH CH.sub.3CO iso-butyl H/H S, S 6 (PL-515)
4-NH.sub.2 3-F 4 (HO).sub.2P(O) (C.sub.6H.sub.5).sub.2CH
CH.sub.3O--CO iso-butyl H/H S, S 7 (PL-521) 4-NH.sub.2 H 4
CH.sub.3CO (C.sub.6H.sub.5).sub.2CH CH.sub.3O--CO iso-butyl H/H S,
S 8 (PL-520) 4-NH.sub.2 H 4 3-Pyridyl-CO (C.sub.6H.sub.5).sub.2CH
CH.sub.3O--CO iso-butyl H/H S, S 9 (PL-534) 4-NH.sub.2 H 4
(CH.sub.3).sub.2NCH.sub.2CO (C.sub.6H.sub.5).sub.2CH CH.sub.3O--CO
iso-butyl H/H S, S 10 (PL-530) 4-NH.sub.2 H 4
(CH.sub.3).sub.2CHCH(NH.sub.2)CO (C.sub.6H.sub.5).sub.2CH
CH.sub.3O--CO iso-butyl H/H S, S 11 (PL-100) 4-NH.sub.2 H 4 H
(C.sub.6H.sub.5).sub.2CH CH.sub.3O--CO iso-butyl H/H 12 (PL-337)
4-NH.sub.2 3-F 4 H C.sub.6H.sub.5).sub.2CH CH.sub.3O--CO iso-butyl
H/H
[0621] To assess the extent of in vivo cleavage of the phosphate
group from the putative compounds, PL-100, PL-462 (based on
PL-100), PL-337 and PL-515 (based on PL-337) compounds were
administered po (per os, (i.e., by mouth)) (50 mg/kg) to male
Sprague-Dawley rats and their plasma concentration measured at
different time intervals post-administration.
[0622] All test articles (PL-100, PL-462, PL-337 and PL-515) were
prepared in different vehicle at the final concentration of 25
mg/mL. The vehicle composition is as follows: (1) 20% ethanol; 50%
propylene glycol; 0.05% w/v Tween 20 and water (Mix); (2) PBS
buffer (PBS).
[0623] Test articles were administered to male Sprague-Dawley rats
at a single oral dose of 50 mg/kg. Each article was tested in three
rats. Blood samples (0.2-0.3 mL) were collected at the post-dose
time of 10, 20, 40, 60, 120, 180 and 360 minutes. The harvested
blood was centrifuged to isolate plasma. The resulting plasma was
separated and stored at -70.degree. C.
[0624] As indicated above, when the Lysine-based compounds
described herein are administered in vivo, the active ingredient is
released and it may therefore inhibit the protease of retroviruses.
For example, as illustrated in FIG. 3 when PL-461 (or PL-462) is
administered in vivo, the phosphate moiety (in the case of PL-461)
is metabolically cleaved to generate the active ingredient; PL-100.
Similarly when PL-515 is administered in vivo, the phosphate moiety
is cleaved to generate PL-337.
[0625] Plasma samples together with standards and quality control
samples were treated to precipitate proteins, then analyzed by
HPLC-MS, for the presence of PL-462, PL-100, PL-515 and PL-337.
TABLE-US-00009 TABLE 11 PL-462 PL-100 PL-515 PL-337 Compound (Ex.
No. 2) (Ex. No. 1-F) (Ex. No. 13) (Ex. No. 13-A) Vehicle PBS Mix
PBS Mix Number of 3 3 3 3 rats Dose (mg/Kg) 50 po 50 po 50 po 50 po
AUC 0.816 .+-. 0.295 0.675 .+-. 0.171 1.075 .+-. 0.625 1.180 .+-.
0.196 (.mu.g/hr * ml) (PL-100, detected) (PL-337, detected) Cmax
(nM) 330 .+-. 109 498 .+-. 203 545 .+-. 215 681 .+-. 131 Tmax (min)
93 .+-. 60 40 .+-. 16 87 .+-. 60 60 .+-. 15 In Table 11 above: 50
mg/Kg PL-462 = 43 mg/Kg PL-100 50 mg/Kg PL-515 = 43 mg/Kg
PL-337
[0626] The results demonstrate that PL-462 and PL-515 compounds may
be delivered orally in aqueous solutions. None of the PL-462 and
PL-515 compounds, delivered as aqueous solutions, are detected in
the blood samples, which suggests rapid metabolism to PL-100 and
PL-337 the parent drugs.
[0627] Aqueous dosing of PL-462 and PL-515 solutions showed
equivalent to slightly superior delivery of PL-100 and PL-337
compared to non-aqueous formulations of PL-100 and PL-337.
[0628] Based on these results, all the phosphorylated compounds
described in the present invention will demonstrate similar
pharmacokinetic properties.
[0629] Partition coefficient (LogP) of selected compounds and the
corresponding HIV protease inhibitors (drug) are as follow:
TABLE-US-00010 TABLE 12 Corresponding Compounds LogP drugs LogP
PL-461 (or PL-462) -1.2 PL-100 3.6 PL-515 -0.75 PL-337 3.8
[0630] The LogP were measured in a standard fashion by dissolving 1
mg of compound in 0.8 mL of each octanol and phosphate buffer pH
7.4 (0.04 M KHPO.sub.4). The concentration of the compounds in the
phases was detected by LC-MS. This test demonstrates the solubility
of the compounds at physiological pH. The LogP obtained show that
the compounds are highly soluble as compared to the corresponding
drugs.
[0631] Results of solubility experiments illustrated at FIG. 4
indicates that PL-461 also possesses a moderate water solubility in
acidic conditions and that the solubility increases dramatically
when pH>4.5. Water solubility of PL-100 and PL-461 at pH 7.5 is
0.079 and 145 mg/ml for PL-100 and PL-461, respectively.
[0632] The phosphorylated version of PL-100; namely PL-461, was
selected for further evaluation due to its stability, solubility
and oral bioavailability.
[0633] Materials and Methods-Pharmacokinetics Studies
[0634] Several studies were conducted in order to test the
pharmacokinetic profile of the Lysine-based compounds described
herein either alone or in combination with CYP450 inhibitors.
[0635] A first study (study 1 #141690) was conducted to evaluate
the pharmacokinetic profile of PL-100 and PL-461 after a single
dose oral administration in combination with ritonavir and two
other studies (study 2 #143656 and study 3 #144536) were conducted
to evaluate the pharmacokinetic profile of PL-461 after a single
dose oral administration in combination with ritonavir.
[0636] Animal tested are Sprague Dawley rats (species Rattus
norvegicus) of 6-7 weeks old at the start of dosing and were
obtained from Charles River (Montreal, Canada). The study usually
comprises about 6 animals per groups.
[0637] Twenty four hours prior to the initiation of the study, rats
were randomly selected into 6 groups, and each group had 6 rats (3
rats for each cage). The rats were fasted 3 h. before dosing and 3
h. after dosing.
[0638] The formulations are prepared as indicated below. The
solutions are stable at room temperature over a week and are
covered with aluminum foil to avoid lights. The mixture contains
Ethanol, Propylene Glycol, Tween 20 and Water for Injection (20%,
50%, 0.1% and 30%, v/v/v/v). The pH of all solutions has been
adjusted to 7.5. The dosing volume for all formulations is 2 mL per
kg body weight.
[0639] Blood sample was collected at 15 min, 30 min, 45 min, 1 h, 2
h, 3 h, 6 h, 12 h, 24 h after the dosing. Blood samples were
centrifuged to separate the plasma and separated plasma samples
were placed into cryovials and frozen at -80.degree. C. for further
analysis. Plasma samples together with standards and quality
control samples were treated by protein precipitation and then
analyzed by HPLC-MS.
[0640] Protein Precipitation
[0641] Briefly, protein precipitation was performed on aliquot of
50.0 mL blank plasma (for standards and quality control samples
(QCs)) or PK plasma. 5.00 .mu.L of spiking solution was added for
standards and QCs or acetonitrile/DMSO=1/1 for PK samples.
[0642] The samples were mixed by vortexing for 10 seconds. 150 uL
of internal standard working solution (1.00 ug/mL PL-459-01 in
acetonitrile) was added. Samples were again vortexed for 10 seconds
and sonicated for 5 mins. The samples were centrifuged for 10 mins
at 10,000 rpm and 100 uL of the supernatant was transferred into an
insert. 100 uL of deionized water (0.2% FA) was added and sample
vortexed. Finally, 20 uL was injected in HPLC. TABLE-US-00011
Instrument Conditions for HPLC HPLC: Column: Zorbax SB-C18, 2.1
.times. 50 mm, 5 .mu.m Mobile phase: aqueous (Aq) = 0.1% formic
acid in deionized water Organic (Org) = 0.1% formic acid in
acetonitrile Gradient: From 30% to 90% of Org in 5 mins, then
return to 30% of Org in 1 min and equilibres 4 mins Flow rate: 0.30
mL/min Run time: 10 mins Retention time: 5.9 min for PL-100 and
PL-459 (IS), 4.6 min for PL-461, 6.3 min for ritonavir MSD: Source:
ESI Peakwidth: 0.07 min Polarity: Positive Acquisition time:
2.0-7.0 mins Acquisition mode: SIM, MH+, 625.3 for PL-100; 630.3
for PL-459 (IS), 705.3 for PL- 461, 721.3 for ritonavir Fragmentor:
100 Gain: 1.5 Resolution: High Spray chamber: Gas temperature:
325.degree. C. Drying gas: 12.0 L/min Nebulizer pressure: 55 psig
Vcap: 4000 V
[0643] Bioavailability Calculations Bioavailability .times. .times.
of .times. .times. PL .times. .times. 461 = ( AUC 0 - 24 .times. h
- p . o . ) ( AUC 0 - 24 .times. h - iv . ) Dose - iv Dose - po M .
W . - PL .times. .times. 461 M . W . - PL .times. .times. 100 100
.times. % ##EQU1## Bioavailability .times. .times. of .times.
.times. PL .times. .times. 100 = ( AUC 0 - 24 .times. .times. h - p
. o . .times. ) ( AUC 0 - 24 .times. .times. h - iv . ) Dose - iv
Dose - po 100 .times. .times. % ##EQU1.2## Note .times. :
##EQU1.3## Molecular .times. .times. Weight .times. .times. of
.times. .times. PL .times. .times. 461 = 705 ##EQU1.4## Molecular
.times. .times. Weight .times. .times. of .times. .times. PL
.times. .times. 100 = 624 ##EQU1.5##
[0644] Test Articles for Studies 1, 2 and 3
[0645] Unless specifically mentioned, female rats were used.
TABLE-US-00012 Test articles of study 1 Name: PL-100-06 Batch/Lot
No.: 06 Storage Conditions: Room Temperature (15-30.degree. C.)
Name: PL-461-05 Batch/Lot No.: 05 (ZL-177-091404) Storage
Conditions: Room Temperature (15-30.degree. C.) Name: Ritonavir
Batch/Lot No.: 749592E22 Storage Conditions: Room Temperature
(15-30.degree. C.)
[0646] Formulation Used in Study 1
[0647] Formulation 1:
[0648] PL-100-06 (30 mg/kg) concentration is 15 mg/mL
[0649] PL-100-06; 90.0 mg dissolved in 6.00 mL Mix
[0650] Formulation 2:
[0651] PL-100-06 (30 mg/kg) concentration is 15 mg/mL and Ritonavir
(10 mg/kg) concentration is 5 mg/mL
[0652] PL-100-06; 90.0 mg+30 mg RTV dissolved in 6.00 mL Mix
[0653] Formulation 3:
[0654] PL-100-06 (30 mg/kg) concentration is 15 mg/mL and Ritonavir
(30 mg/kg) concentration is 15 mg/mL
[0655] PL-100-06; 90.0 mg+90 mg RTV dissolved in 6.00 mL Mix
[0656] Formulation 4:
[0657] PL-461-05 (30 mg/kg) concentration is 15 mg/mL
[0658] PL-461-05; 90.0 mg dissolved in 6.00 mL Mix, pH 7.0
[0659] Formulation 5:
[0660] PL-461-05 (30 mg/kg) concentration is 15 mg/mL and Ritonavir
(10 mg/kg) concentration is 5 mg/mL
[0661] PL-461-05; 90.0 mg+30 mg RTV dissolved in 6.00 mL Mix, pH
7.0
[0662] Formulation 6:
[0663] PL-461-05 (30 mg/kg) concentration is 15 mg/mL and Ritonavir
(30 mg/kg) concentration is 15 mg/mL
[0664] PL-461-05; 90.0 mg+90 mg RTV dissolved in 6.00 mL Mix, pH
7.0 TABLE-US-00013 Test articles of study 2 Name: PL-461-05
Batch/Lot No.: 05 (ZL-7-177-091404) Storage Conditions: Room
Temperature (15-30.degree. C.) Name: Ritonavir Batch/Lot No.:
749592E22 Storage Conditions: Room Temperature (15-30.degree.
C.)
[0665] Formulation Used in Study 2
[0666] Formulation 1:
[0667] PL-461-05 (29.93 mg/kg) concentration is 14.97 mg/mL
[0668] PL-461-05; 89.8 mg dissolved in 6.00 mL Mix
[0669] Formulation 2:
[0670] PL-461-05 (30.2 mg/kg) concentration is 15.10 mg/mL and
Ritonavir (10 mg/kg) concentration is 5 mg/mL
[0671] PL-461-05; 90.6 mg+30 mg RTV dissolved in 6.00 mL Mix
[0672] Formulation 3:
[0673] PL-461-05 (29.43 mg/kg) concentration is 14.72 mg/mL and
Ritonavir (5.10 mg/kg) concentration is 2.55 mg/mL
[0674] PL-461-05; 88.3 mg+15.3 mg RTV dissolved in 6.00 mL Mix
[0675] Formulation 4:
[0676] PL-461-05 (51.27 mg/kg) concentration is 25.63 mg/mL
[0677] PL-461-05; 153.8 mg dissolved in 6.00 mL Mix, pH 7.0
[0678] Formulation 5:
[0679] PL-461-05 (50.03 mg/kg) concentration is 25.02 mg/mL and
Ritonavir (17.03 mg/kg) concentration is 8.52 mg/mL
[0680] PL-461-05; 150.1 mg+51.1 mg RTV dissolved in 6.00 mL Mix, pH
7.0
[0681] Formulation 6:
[0682] PL-461-05 (49.33 mg/kg) concentration is 24.67 mg/mL and
Ritonavir (8.03 mg/kg) concentration is 4.02 mg/mL
[0683] PL-461-05; 148.0 mg+24.1 mg RTV dissolved in 6.00 mL Mix, pH
7.0 TABLE-US-00014 Test articles of study 3 Name: PL-461-06
Batch/Lot No.: 06 (SAP-8-57-102604) Storage Conditions: Room
Temperature (15-30.degree. C.) Name: Ritonavir (RTV) Batch/Lot No.:
749592E22 Storage Conditions: Room Temperature (15-30.degree.
C.)
[0684] Formulation Used in Study 3
[0685] Formulation 1:
[0686] PL-461-06 (30 mg/mL) dose is 60 mg/kg
[0687] PL-461-06; 180.0 mg dissolved in 6.00 mL Mix
[0688] Formulation 2:
[0689] PL-461-06 (15 mg/mL) dose is 30 mg/kg and Ritonavir (5
mg/mL) dose is 10 mg/kg
[0690] PL-461-06; 90.0 mg+30 mg RTV dissolved in 6.00 mL Mix
[0691] Formulation 3:
[0692] PL-461-06 (50 mg/mL) dose is 100 mg/kg and Ritonavir (5
mg/mL) dose is 10 mg/kg
[0693] PL-461-06; 300.0 mg+30 mg RTV dissolved in 6.00 mL Mix
[0694] Formulation 4:
[0695] PL-461-06 (50 mg/mL) dose is 100 mg/kg
[0696] PL-461-06; 300.0 mg dissolved in 6.00 mL Mix
[0697] Formulation 5:
[0698] PL-461-06 (30 mg/mL) dose is 60 mg/kg and Ritonavir (5
mg/mL) dose is 10 mg/kg
[0699] PL-461-06; 180.0 mg+30 mg RTV dissolved in 6.00 mL Mix
[0700] Formulation 6:
[0701] PL-461-06 (50 mg/mL) dose is 100 mg/kg and Ritonavir (8.33
mg/mL) dose is 16.67 mg/kg
[0702] PL-461-06; 300 mg+50 mg RTV dissolved in 6.00 mL Mix
TABLE-US-00015 Test articles of study 4 PL-461-06 Batch/Lot No.: 06
(SAP-8-57-102604) Storage Conditions: Room Temperature
(15-30.degree. C.) Ritonavir (RTV) Batch/Lot No.: 21-802AW21
Storage Conditions: Room Temperature (15-30.degree. C.)
Manufacturer: Abbott Laboratories, Limited
[0703] TABLE-US-00016 Preparation used in study 4 PL-461-06
Preparation 1 Test Articles: PL-461 (20 mg/mL) Vehicle: Water for
injection pH: 6.0 Appearance: Pale yellow solution Storage
Condition: Room temperature (15-30.degree. C.) PL-461-06
Preparation 2 Test Articles: PL-461 (60 mg/mL) Vehicle: Water for
injection pH: 6.0 Appearance: Pale yellow solution Storage
Condition: Room temperature (15-30.degree. C.) PL-461-06
Preparation 3 Test Articles: PL-461 (200 mg/mL) Vehicle: Water for
injection pH: 6.0 Appearance: Yellow solution Storage Condition:
Room temperature (15-30.degree. C.)
[0704] Ritonavir Solution (16 mq/mL)
[0705] 2 mL of the Ritonavir Solution, Norvir, (80 mg/mL) is
diluted with 8 mL mixture containing 60% Propylene Glycol and 40%
Water. The final concentration of Ritonavir is 16.0 mg/mL.
Example 14
Pharmacokinetics of PL-461
[0706] The pharmacokinetics (PK) of PL-461 was tested and compared
with that of the original active ingredient.
[0707] The PK profile was obtained by administration of the test
drugs; PL-100 and PL-461 in rats. PL-100 or PL-461 was administered
orally at doses indicated in FIG. 5. Each time point in the figure
represents the average plasma [PL-100] (ng/ml) of 6 female rats at
a given dose.
[0708] The results indicate that the absolute oral bioavailability
for PL-100 and PL-461 at 30 mg/kg is 8.7 and 23%, respectively. The
bioavailability of PL-461 at 100 mg/kg is 23%.
[0709] The horizontal line in FIG. 5 represents plasma [PL-100] of
200 ng/ml. The PK profile of PL-461 shows that the time (t)>200
ng/ml is approximately 6 hours in rats at 100 mg/kg, suggesting
PL-461 has a potential as a twice daily drug to maintain such
plasma [PL-100] in man at an equivalent dose.
[0710] The pharmacokinetic parameters of PL-100 and PL-461 in
rats** was compared. These results are summarized in Table 13
below. TABLE-US-00017 TABLE 13 Dose .lamda.z T1/2 Tmax Cmax Tlast
Clast AUClast AUCinf Vz_F Cl_F mg/kg Route (1/hr) (hr) (hr) (ng/ml)
(hr) (ng/ml) (hr ng/ml) (hr ng/ml) (L/kg) (L/hr/kg) PL-100* 5 i.v.
0.52 1.2 0.1 1804 6 44 1369 1434 6.1 3.5 PL-100* 50 p.o. 0.09 8.4
0.4 457 24 15 1101 1284 549 44 PL-461* 50 p.o. 0.10 9.4 0.5 1216 24
13 2226 2416 282 23 In Table 13, the abbreviations are as follows:
.lamda.z elimination rate constant t1/2 plasma elimination
half-life Tmax time of maximum concentration Cmax maximum
concentration Tlast time of last measurable concentration Clast
last measurable concentration AUClast area under the plasma
conc-time curve from zero to the last measurable concentration
AUCinf area under the plasma concentration-time curve from zero
extrapolated to infinity Vz_F apparent volume of distribution Cl_F
apparent oral clearance *Each treatment group had at least 6 female
rats. **PK analysis was done using the non-compartmental method
with WinNonLin Professional (version 4.0). AUC was calculated using
the linear-up/log-down method. Results presented in FIG. 5 and in
Table 13 indicate that PL-461 has improved PK over PL- 100.
Example 15
Methods and Compositions for Improving the Pharmacokinetics of
Active Ingredients and Lysine-Based Compounds
[0711] The results presented herein indicate that although the
active ingredients (e.g., PL-100, etc.) and the Lysine-based
compounds (e.g., PL-461, PL-462, etc.) are good anti-viral
compounds, the pharmacokinetics may still be improved.
[0712] Further experiments were therefore conducted to that effect
by administering PL-100 or PL-461 with a drug able to inhibit
cytochrome P450 monooxygenase (CYP450) (i.e., a CYP450 inhibitor)
in order to test whether this type of inhibitor may increase the
pharmacokinetics of compounds described herein.
[0713] Ritonavir, quinidine, ketoconazole and sulfaphenazole were
chosen among candidate CYP450 inhibitors and the pharmacokinetics
of PL-461 and CYP450 inhibitors combinations was compared with the
pharmacokinetics of other PI and CYP450 inhibitors combination. The
effect of CYP450 inhibitors on the metabolism of HIV-1
anti-protease was tested in human liver microsomes.
[0714] Although, ketoconazole and sulfaphenazole are able to
increase the PK of PL-100, results of FIG. 6 indicate that the most
effective combination is that of PL-100 and ritonavir.
[0715] Results presented in FIG. 7 and in Table 14 illustrate the
bioavailability of PL-100 or PL-461 when administered in
combination with a CYP450 inhibitor. TABLE-US-00018 TABLE 14 Oral
bioavailability of PL-100 and PL-461 in female rats (# 141690)
Group Test Dose* Dose** No. article (mg/Kg) (mg/Kg) Absolute
Bioavailability*** (%) Ave. SD CV (%) Max Min Median 1 PL100/RTV
30/0 30/0 8.5 8.4 8.5 9.0 7.2 10.7 8.7 1.1 12.8 10.7 7.2 8.5 2
30/10 33/10 36.3 25.0 24.8 31.3 21.9 33.7 28.8 6.3 19.9 36.3 21.9
28.1 3 30/30 32/30 70.0 54.6 75.3 54.8 70.2 41.2 61.0 13.0 21.3
75.3 41.2 62.4 4 PL461/RTV 30/0 35/0 15.9 33.6 13.0 14.9 22.3 23.8
20.6 7.7 37.3 33.6 13.0 19.1 5 30/10 34/11 80.7 45.1 105.1 94.3
47.7 36.5 68.2 28.8 42.3 105.1 36.5 64.2 6 30/30 34/34 32.7 93.8
42.1 64.9 66.7 43.5 57.3 22.4 39.1 93.8 32.7 54.2 *Intended dose by
protocol **Corrected dose by QC results. ***Bioavailability was
calculated from corrected dose.
[0716] These results indicate that ritonavir (RTV: a CYP450
inhibitor) increases the bioavailability of PL-100 and that of
PL-461 as well.
[0717] Further CYP450 inhibitor efficient at increasing the
pharmacokinetics of the active ingredients or the Lysine-based
compounds (referred here as "compounds") described herein may be
identified by co-administering a putative CYP450 inhibitor and a
desired compound (one or more compounds) in an assay (e.g.,
microsome assay or in vivo studies) described herein and to measure
the amount of residual compound (active ingredient) still remaining
after various time points following administration (incubation). A
CYP450 inhibitor which increases the bioavailability of the
compound (active ingredient) compared to the bioavailability
measured without the putative CYP450 inhibitor would be found
efficient for the purpose of improving the pharmacokinetics of the
compound.
[0718] In study 4, ritonavir was first administered and PL-1461 was
administered 15 or 30 minutes later. The result obtained using this
administration scheme (Table 15) indicates that this method is as
efficient at increasing the pharmacokinetics of the Lysine-based
compound disclosed herein as the method of studies 1-3.
TABLE-US-00019 TABLE 15 Conc. of Test Dosing Number of Article
Route of Volume Animals per Group Test Article Dose (mg/kg) (mg/mL)
Administration (mL/kg) Group 1 PL-461-06 100 20 Oral 5 3 male/ 3
female 2 PL-461-06 300 60 Oral 5 3 male/ 3 female 3 PL-461-06 1000
200 Oral 5 3 male/ 3 female 4 RTV 16 16 Oral 1 3 male/ PL-461-06
100 20 Oral 5 3 female 5 RTV 16 16 Oral 1 3 male/ PL-461-06 100 20
Oral 5 3 female 6 RTV 16 16 Oral 1 3 male/ PL-461-06 100 20 Oral 5
3 female
Example 16
Pharmacokinetics of Lysine-Based Compounds and CYP450 Inhibitors
Combination
[0719] In studies 1, 2 and 3 mentioned above, PL-461 and RTV were
co-administered orally at doses indicated in FIG. 8.
[0720] Each time point in FIG. 8 represents the average plasma
concentration of PL-100 (ng/ml) of 6 female rats at a given dose.
The horizontal line in FIG. 8 represents plasma concentration of
PL-100 of 630 ng/ml. The PK profile shows that the time (t)>630
ng/ml is approximately 6 hours in rats at 100 mg/kg PL-461 and 16.7
mg/kg RTV, suggesting that PL-461 has a potential as a twice daily
drug to maintain such plasma concentration of PL-100 in man at an
equivalent dose.
Example 17
Optimization of Lysine-Based Compounds vs CYP450 Inhibitor
Ratio
[0721] The absolute oral bioavailability of PL-461 was determined
under various conditions indicated in FIG. 9. More particularly,
the concentration of PL-461 and the proportion of PL-461 compared
to CYP450 was varied. Results presented herein indicate that
sufficient oral bioavailability, relative to protein binding
adjusted EC.sub.95 against resistant strains may be achieved when
boosted at a ratio of 6 (PL-461) to 1 (RTV), although other ratios
may also successfully be used as indicated in Table 16 below.
TABLE-US-00020 TABLE 16 Oral bioavailability of PL-461 in rats with
and without ritonavir (#141690, #143656, #144536) Dose level Ratio
of dose Dose* (mg/kg) of PL461 (PL461/RTV) (PL461/RTV) Absolute
Bioavailability (%)** Ave. SD CV (%) Max Min Median 30 mg/kg N/A***
35/0 15.9 33.6 13.0 14.9 22.3 23.8 22.5 6.9 30.7 33.6 13.0 23.8
27/0 30.1 24.0 24.5 6/1 26/3.5 42.2 37.6 45.1 65.5 47.4 12.3 26.0
65.5 37.6 43.7 3/1 34/11 80.7 45.1 105.1 94.3 47.7 36.5 52.6 24.5
46.6 105.1 26.5 43.9 26/9.2 62.1 45.3 26.5 38.5 42.6 34.7 26/8.4
94.7 37.1 32.9 28.6 54.9 39.4 1/1 34/34 32.7 93.8 42.1 64.9 66.7
43.5 57.3 22.4 39.1 93.8 32.7 54.2 50 mg/kg N/A*** 48/0 11.1 20.3
9.2 23.3 31.3 17.4 7.7 44.6 31.3 7.1 14.7 51/0 12.0 7.1 14.7 24.4
13.4 24.4 6/1 55/11 34.9 51.7 33.9 58.4 46.0 24.1 41.5 12.8 30.7
58.4 24.1 40.5 3/1 45/15 49.0 64.5 41.9 42.3 13.8 85.2 49.5 24.0
48.6 85.2 13.8 45.7 100 mg/Kg N/A*** 84/0 13.6 37.1 25.3 25.6 15.2
23.4 9.5 40.6 37.1 13.6 25.3 10/1 85/9.2 25.2 20.1 40.0 20.3 28.9
51.4 31.0 12.4 40.0 51.4 20.1 27.1 6/1 89/17 91.2 47.0 39.1 56.7
35.6 53.9 22.4 41.5 91.2 35.6 47.0 *Corrected dose by QC results.
**Bioavailability was calculated by corrected dose. ***No
Ritonavir
[0722] Finally FIG. 10 indicates that PL-100 inhibits CYP3A4/5 as
does RTV. RTV's higher Ki (about 10-fold higher) confirms that it
is able to act as a boosting agent for PL-100.
[0723] It was shown herein that PL-100 is a potent, specific and
non-cytotoxic novel PI and that it has a favorable cross-resistance
pattern compared to all approved Pls.
[0724] PL-461, a precursor of PL-100, is >1800-fold more water
soluble than PL-100 and has a 2 to 3-fold improved oral
bioavailability over PL-100. PL-461 has a potential as a novel PI
for the treatment of patients infected with PI-resistant HIV
strains bearing, for example, two or less primary mutations.
PL-461, when combined with a CYP450 inhibitor such as, for example,
ritonavir, has a great potential as a novel PI for the treatment of
patients infected with PI-resistant HIV strains bearing mutations
(e.g., two or more primary mutations). The ratio of PL-461 to
ritonavir required for boosting has been found herein adequate for
in vivo administration.
[0725] On the basis of PL-100 cross-resistance data and the
pharmacokinetics results, it is expected that equivalent doses of
PL-461 in man will effectively inhibit replication of
protease-resistant HIV strains. Moreover, these data also suggests
that PL-461 may be administered, for example, using a convenient
twice-daily dosing regimen.
[0726] It is understood that the examples and embodiments described
herein are for illustrative purposes only and that various
modifications or changes in light thereof will be suggested to
persons skilled in the art and are to be included within the spirit
and purview of this application and scope of the appended claims.
All publications, patents, and patent applications cited herein are
hereby incorporated by reference in their entirety for all
purposes.
* * * * *